Peptidic procollagen C-proteinase inhibitors

ABSTRACT

This invention relates to compounds of Formula (I)  
                 
 
     wherein R 1 -R 7 , A, Z and n are as described in the Summary of the Invention that are inhibitors of procollagen C-proteinase, pharmaceutical compositions containing them, methods for their use and methods for their preparation.

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. 119(e) ofU.S. Provisional Application Serial No. 60/111,661, filed Dec. 10, 1998,hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to compounds that inhibitprocollagen C-proteinase, pharmaceutical compositions containing them,methods for their use, and methods for preparing these compounds.

BACKGROUND INFORMATION AND RELATED DISCLOSURES

[0003] The collagens are integral components of connective tissue. Atpresent nineteen types of collagens have been identified. Theinterstitial collagen types I, II and III are the major collagencomponents of tissue. These collagens are synthesized as procollagenprecusor molecules having amino- and carboxy-terminal peptide extensionsalso known as pro-regions. These pro-regions are typically cleaved uponsecretion of the procollagen molecule to give a mature collagen moleculewhich is capable of association into highly structured collagen fibers.(see, e.g., Fessler and Fessler, Annu. Rev. Biochem. 47, 129, (1978);Kivirikko et al., Extracellular Matrix Biochemistry (1984) and Kuhn,Structure and Function of Collagen Types (eds Mayne, R and Burgeson, R.E.), Academic Press, Inc., Orlando, Fla. pp 1-42 (1987).

[0004] Excessive collagen deposition is associated with a variety offibrotic diseases such as interstitial pulmonary fibrosis, pericentralfibrosis, Symmers' fibrosis, perimuscular fibrosis, kidney fibrosis,endocardial sclerosis, hepatitis, acute respiratory distress syndrome,arthritis, cystic fibrosis, surgical adhesions, tendon surgery, cornealscarring, scleroderma, chronic allograft rejection, hemodialysis shuntfibrosis and restenosis. These diseases are characterized by excessivedeposits of fibrillar interstitial collagens that are resistant toproteolyic degradation thus leading to the symptoms of fibrosis.Therefore, inhibition of the pathological deposition of thse collagensshould help in the treatment of these diseases.

[0005] Recent studies suggest that procollagen C-proteinase is theessential enzyme that catalyzes the cleavage of the C-propeptide oftypes I, II and III collagens and therefore instrumental in theformation of functional collagen fibers ((see, Fertala et al., J. Biol.Chem., 269, 11584, (1994)). It would therefore be desirable to provideprocollagen C-proteinase inhibitors and thereby provide a means ofcombating diseases mediated by excessive deposition of these collagens.This invention fulfills this and related needs.

SUMMARY OF THE INVENTION

[0006] In a first aspect, this invention provides hydroxamic acidsselected from the group of compounds represented by Formula (I):

[0007] wherein:

[0008] R¹ and R⁴ are, independently of each other, hydrogen or alkyl;

[0009] R² is:

[0010] (i) cycloalkyl, cycloalkylalkyl, aryl, aralkyl, aralkenyl,heteroaryl, heteroaralkyl, heteroaralkenyl, heterocyclo orheterocycloalkyl; or

[0011] (ii) -(alkylene)-B—X where B is —O—, —NR⁸—, —S(O)_(n)— (where nis 0, 1 or 2), —C═O, —CONR⁸—, —NR⁸CO₂—, NR⁸SO₂or —C(═NR⁸)NR⁸SO₂— (whereR⁸is H or alkyl), and X is cycloalkyl, cycloalkylalkyl, aryl, aralkylheteroaryl or heteroaralkyl; or

[0012] (iii) -(alkylene)-B—X where B is —NR⁸CO— (where R⁸ is H oralkyl), and X is cycloalkyl, cycloalkylalkyl, aryl, aralkyl heteroarylor heteroaralkyl; or

[0013] (iv) R² and R³ form an alkylene or heteroalkylene chain;

[0014] R³ is hydrogen or alkyl;

[0015] R⁶ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl,aralkyl, heteroaryl or heteroaralkyl;

[0016] R⁵ is:

[0017] (i) hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl,aralkenyl, heteroaryl, heteroaralkyl, heteroaralkenyl, heterocycloalkyl,heteroalkyl, or -(alkylene)-C(O)—X¹ where X¹ is alkyl, hydroxy, alkoxy,aryl, aralkyl, aryloxy, aralkyloxy, heteroaryl, heteroaryloxy,heteroaralkyloxy or NR′R″ (where R′ and R″ are independently H or alkyl,or R′ and R″ form an alkylene chain); or

[0018] (ii) R⁵ and R⁴ form an alkylene chain; or

[0019] (iii) R⁵ and R⁶ form an alkylene chain;

[0020] n is 0 or 1;

[0021] A is —C(═O)—CH(R⁹)—(CH₂)_(m)—N(R¹⁰)— wherein:

[0022] m is an integer from 0-5 inclusive;

[0023] R⁹ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl,aralkyl, heteroaryl, heteroaralkyl, heterocycloalkyl, heteroalkyl, or-(alkylene)-C(O)—X¹ where X¹ is alkyl, hydroxy, alkoxy, aryl, aralkyl,aryloxy, aralkyloxy, heteroaryl, heteroaryloxy, heteroaralkyloxy orNR′R″ (where R′ and R″ are independently H or alkyl, or R′ and R″ forman alkylene chain); and

[0024] R¹⁰ is hydrogen, alkyl, aralkyl or heteroaralkyl;

[0025] Z is Y—B wherein:

[0026] Y is alkylene or a bond; and

[0027] B is —CO—, —C(O)O—, —CONR⁸—, —SO₂—, or —SO₂NR⁸— (where R⁸ ishydrogen or alkyl), alkylene (optionally substituted by hydroxy, alkoxy,amino, monoalkylamino or dialkylamino) or a bond;

[0028] R⁷ is cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl orheteroaralkyl;

[0029] provided that when n=0 and Z is SO₂, then R² does not contain animidazole group; and their pharmaceutically acceptable salts, prodrugs,individual isomers, and mixtures of isomers.

[0030] In a second aspect, this invention provides a method of treatmentof a disease in a mammal treatable by administration of a procollagenC-proteinase inhibitor selected from the group of compounds representedby Formula (I):

[0031] In a third aspect, this invention provides pharmaceuticalcompositions containing a therapeutically effective amount of a compoundof Formula (I) or its pharmaceutically acceptable salt and apharmaceutically acceptable excipient.

[0032] In a fourth aspect, this invention provides a method of treatingdisease by administering to a patient a selective inhibitor ofprocollagen-C-proteinase.

[0033] In a fifth aspect, this invention provides a method of preparingcompounds of Formula (I).

DETAILED DESCRIPTION OF THE INVENTION Definitions

[0034] Unless otherwise stated, the following terms used in thespecification and claims have the meanings given below:

[0035] “Alkyl” means a linear saturated monovalent hydrocarbon radicalof one to six carbon atoms or a branched saturated monovalenthydrocarbon radical of three to six carbon atoms, e.g., methyl, ethyl,propyl, 2-propyl, pentyl, and the like.

[0036] “Alkylene” means a linear saturated divalent hydrocarbon radicalof one to six carbon atoms or a branched saturated divalent hydrocarbonradical of three to six carbon atoms, e.g., methylene, ethylene,propylene, 2-methylpropylene, pentylene, and the like.

[0037] “Heteroalkylene” means an alkylene chain in which one methylenegroup has been replaced by O, S or NR′ (where R′ is hydrogen or alkyl.)

[0038] “Alkenyl” means a linear monovalent hydrocarbon radical of two tosix carbon atoms or a branched monovalent hydrocarbon radical of threeto six carbon atoms, containing at least one double bond, e.g., ethenyl,propenyl, and the like.

[0039] “Alkenylene” means a linear divalent hydrocarbon radical of twoto six carbon atoms or a branched monovalent hydrocarbon radical ofthree to six carbon atoms, containing at least one double bond, e.g.,ethenylene, 2-propenylene, and the like.

[0040] “Acyl” means a radical —C(O)R where R is hydrogen, alkyl,alkenyl, cycloalkyl, cycloalkylalkyl, haloalkyl, aryl, aralkyl,aralkenyl, heteroaryl, heteroaralkyl, heteroaralkenyl, or heterocyclo,e.g., acetyl, benzoyl, thenoyl, and the like.

[0041] “Acyloxy” means a radical —OC(O)R where R is hydrogen, alkyl,alkenyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, aralkenyl, orhaloalkyl, e.g., acetoxy, 3,3,3-trifluoroacetoxy and the like.

[0042] “Acylamino” means a radical —NRC(O)R′ where R is hydrogen oralkyl and R′ is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,heteroalkyl, haloalkyl, aryl, aralkyl, aralkenyl, heteroaryl,heteroaralkenyl, or heteroaralkyl, e.g., acetylamino,trifluoroacetylamino, benzoylamino, methylacetylamino, and the like.

[0043] “Sulfonylamino” means a radical —NRSO₂R′ where R is hydrogen oralkyl and R′ is alkyl, alkenyl, cycloalkyl, cycloalkylalkyl,heteroalkyl, haloalkyl, amino, monosubstituted amino, disubstitutedamino, aryl, aralkyl, aralkenyl, heteroaryl, heteroaralkenyl, orheteroaralkyl, e.g., methylsulfonylamino, benzylsulfonylamino,N-methylaminosulfonylamino, and the like.

[0044] “Halo” means fluoro, chloro, bromo, or iodo, preferably fluoroand chloro.

[0045] “Haloalkyl” means alkyl substituted with one or more same ordifferent halo atoms, e.g., —CH₂Cl, —CF₃, —CH₂CF₃, —CH₂CCl₃, and thelike.

[0046] “Cycloalkyl” means a saturated monovalent cyclic hydrocarbonradical of three to six ring carbons, e.g., cyclopropyl, cyclopentyl,cyclohexyl, and the like.

[0047] “Carbocycle” means a saturated, cyclic group of 3 to 8 ring atomsin which all the ring atoms are carbon, e.g., cyclopentyl, cyclohexyl,and the like.

[0048] “Aryl” means a monovalent monocyclic or bicyclic aromatichydrocarbon radical of 6 to 10 ring atoms, optionally fused to acarbocycle or heterocycle, and optionally substituted independently withone or more substituents, preferably one or two substituents selectedfrom alkyl, heteroalkyl, haloalkyl, halo, nitro, acyloxy, cyano,cycloalkyl, cycloalkylalkyl, optionally substituted phenyl, optionallysubstituted phenylalkyl, heteroaryl, heteroaralkyl, —OR (where R ishydrogen, alkyl, haloalkyl, alkenyl, cycloalkyl, cycloalkylalkyl,optionally substituted phenyl, heteroaryl, optionally substitutedphenylalkyl, or heteroaralkyl), —NRR′ (where R and R′ are independentlyhydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl,optionally substituted phenyl, optionally substituted phenylalkyl,optionally substituted phenylalkenyl, heteroaryl, or heteroaralkyl),—C(O)R (where R is hydrogen, alkyl, alkenyl, cycloalkyl,cycloalkylalkyl, haloalkyl, optionally substituted phenyl, optionallysubstituted phenylalkyl, optionally substituted phenylalkenyl,heteroaryl, heteroaralkyl, or heteroaralkenyl), —S(O)_(n)R (where n isan integer from 0 to 2 and R is hydrogen (provided that n is 0), alkyl,haloalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, optionally substitutedphenyl, heteroaryl, optionally substituted phenylalkyl, orheteroaralkyl), —SO₂NRR′ (where R and R′ are independently hydrogen,alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, optionallysubstituted phenyl, optionally substituted phenylalkyl, optionallysubstituted phenylalkenyl, heteroaryl or heteroaralkyl, or R and R′together with the nitrogen they are attached to form a cycloamino ring),—COOH, -(alkylene)-COOH, -(alkenylene)-COOH, —COOR^(a),-(alkenylene)-COOR^(a), -(alkylene)-COOR^(a) (where R^(a) is alkyl,optionally substituted phenylalkyl, or heteroaralkyl), —CONR′R″,-(alkylene)-CONR′R″, (where R′ and R″ are independently selected fromhydrogen, alkyl, cycloalkyl, cycloalkylalkyl, optionally substitutedphenyl, optionally substituted phenylalkyl, heteroaryl andheteroaralkyl, or R′ and R″ together with the nitrogen they are attachedto form a cycloamino ring), —NRC(O)R′ (where R is hydrogen or alkyl andR′ is hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, haloalkyl,optionally substituted phenyl, optionally substituted phenylalkyl,optionally substituted phenylalkenyl, heteroaryl, heteroaralkenyl, orheteroaralkyl), —NRSO₂R′ (where R is hydrogen or alkyl and R′ is alkyl,alkenyl, cycloalkyl, cycloalkylalkyl, haloalkyl, optionally substitutedphenyl, optionally substituted phenylalkyl, optionally substitutedphenylalkenyl, heteroaryl, heteroaralkenyl, or heteroaralkyl), or—NRSO₂NR′R″ (where R is hydrogen or alkyl and R′ and R″ areindependently hydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl,cycloalkylalkyl, optionally substituted phenyl, optionally substitutedphenylalkyl or optionally substituted phenylalkenyl, or R′ and R″together with the nitrogen they are attached to form a cycloamino ring).More specifically the term aryl includes, but is not limited to, phenyl,1-naphthyl, 2-naphthyl, tetrahydronaphthyl, methylenedioxyphenyl,indanyl, tetralyl, indolinyl, chromanyl, isochromanyl and the like.

[0049] “Heteroaryl” means a monovalent monocyclic or bicyclic aromaticradical of 5 to 10 ring atoms containing one, two, or three ringheteroatoms selected from N, O, or S, the remaining ring atoms being C.The heteroaryl ring is optionally substituted independently with one ormore substituents, preferably one or two substituents, selected fromalkyl, haloalkyl, halo, nitro, cyano, cycloalkyl, cycloalkylalkyl,optionally substituted phenyl, optionally substituted phenylalkyl, —OR(where R is hydrogen, alkyl, haloalkyl, alkenyl, cycloalkyl,cycloalkylalkyl, optionally substituted phenyl or optionally substitutedphenylalkyl), —NRR′ (where R and R′ are independently hydrogen, alkyl,alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, optionally substitutedphenyl, optionally substituted phenylalkyl or optionally substitutedphenylalkenyl, or R and R″ together with the nitrogen they are attachedto form a cycloaraino ring), —C(O)R (where R is hydrogen, alkyl,alkenyl, cycloalkyl, cycloalkylalkyl, haloalkyl, optionally substitutedphenyl, optionally substituted phenylalkyl, or optionally substitutedphenylalkenyl), —S(O)_(n)R (where n is an integer from 0 to 2 and R ishydrogen (provided that n is 0), alkyl, haloalkyl, alkenyl, cycloalkyl,cycloalkylalkyl, optionally substituted phenyl, or optionallysubstituted phenylalkyl), —SO₂NRR′ (where R and R′ are independentlyhydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl,optionally substituted phenyl, optionally substituted phenylalkyl oroptionally substituted phenylalkenyl, or R and R″ together with thenitrogen they are attached to form a cycloamino ring), —COOH,-(alkylene)-COOH,-(alkenylene)COOH, —COOR^(a), -(alkenylene)-COOR^(a),-(alkylene)-COOR^(a) (where R^(a) is alkyl, or optionally substitutedphenylalkyl), —CONR′R″, -(alkylene)-CONR′R″, (where R′ and R″ areindependently selected from hydrogen, alkyl, cycloalkyl,cycloalkylalkyl, optionally substituted phenyl and optionallysubstituted phenylalkyl, or R′ and R″ together with the nitrogen theyare attached to form a cycloamino ring), —NRC(O)R′ (where R is hydrogenor alkyl and R′ is hydrogen, alkyl, alkenyl, cycloalkyl,cycloalkylalkyl, haloalkyl, optionally substituted phenyl, optionallysubstituted phenylalkyl, or optionally substituted phenylalkenyl),—NRSO₂R′ (where R is hydrogen or alkyl and R′ is alkyl, alkenyl,cycloalkyl, cycloalkylalkyl, haloalkyl, optionally substituted phenyl,optionally substituted phenylalkyl, or optionally substitutedphenylalkenyl), —NRSO₂NR′R″ (where R is hydrogen or alkyl and R′ and R″are independently hydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl,cycloalkylalkyl, optionally substituted phenyl, optionally substitutedphenylalkyl or optionally substituted phenylalkenyl, or R′ and R″together with the nitrogen they are attached to form a cycloamino ring),or an amino protecting group. More specifically the term heteroarylincludes, but is not limited to, furyl, thienyl, pyrroly, pyridyl,purinyl, pyrimidinyl, pyrazolyl, thiazolyl, imidazolyl, thiazolyl,thiadiazolyl, indolyl, azaindolyl, benzofuranyl, benzimidazolyl,benzthiazolyl, quinolinyl, isoquinolinyl, benzotriazolyl, benzopyranyl,1, and the derivatives thereof.

[0050] “Optionally substituted phenyl” means phenyl ring which isoptionally substituted with one or more substituents, preferably one ortwo substituents selected from alkyl, haloalkyl, halo, nitro, cyano,—NRR′ (where R and R′ are independently selected from hydrogen andalkyl, or R and R′ together with the nitrogen they are attached to forma cycloamino ring), —OR (where R is hydrogen, alkyl or haloalkyl),—COOR^(a) (where R^(a) is hydrogen or alkyl) or —CONR′R″ (where R′ andR″ are independently selected from hydrogen and alkyl, or R′ and R″together with the nitrogen they are attached to form a cycloamino ring).Representative examples include, but are not limited to, 4-fluorophenyl,3,4-dibromophenyl, 4-chloro-2,5-dimethylphenyl, 2,4,5-trichlorophenyl,4-bromo-2-trifluoromethoxyphenyl, 2-chloro-4-trifluoromethyl,4-tertbutylphenyl, 4-methoxyphenyl, 3-nitrophenyl, and the like.

[0051] “Heterocycle” or “Heterocyclo” means a saturated cyclic radicalof 3 to 8 ring atoms in which one or two ring atoms are heteroatomsselected from N, O, or S(O). (where n is an integer from 0 to 2), theremaining ring atoms being C, where one or two C atoms may optionally bereplaced by a carbonyl group. The heterocyclo ring may be optionallysubstituted independently with one, two, or three substituents selectedfrom alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl,heteroaryl, heteroaralkyl, halo, cyano, acylamino, amino,monosubstituted amino, disubstituted amino, —OR (where R is hydrogen,alkyl, haloalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, aryl,heteroaryl, aralkyl, or heteroaralkyl), —C(O)R (where R is hydrogen,alkyl, alkenyl, cycloalkyl, cycloalkylalkyl, haloalkyl, aryl, aralkyl,aralkenyl, heteroaryl, heteroaralkyl, or heteroaralkenyl), —S(O)_(n)R[where n is an integer from 0 to 2 and R is hydrogen (provided that n is0), alkyl, haloalkyl, alkenyl, cycloalkyl, cycloalkylalkyl, amino,monosubstituted amino, disubstituted amino, aryl, heteroaryl, aralkyl,or heteroaralkyl], —COOH, -(alkylene)-COOH, —COOR^(a),-(alkylene)-COOR^(a) (where R^(a) is alkyl, heteroalkyl, aralkyl, orheteroaralkyl), —CONR′R″, -(alkylene)-CONR′R″ (where R′ and R″ areindependently selected from hydrogen, alkyl, cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heteroaryl and heteroaralkyl, or R′ andR″ together with the nitrogen they are attached to form a cycloaminoring) or an amino protecting group. More specifically the termheterocyclo includes, but is not limited to, tetrahydropyranyl,piperidino, piperazino, morpholino and thiomorpholino,thiomorpholino-1-oxide, thiomorpholino-1,1-dioxide, and the derivativesthereof.

[0052] The term “cycloamino” means a heterocyclo group in which at leastone ring atom is nitrogen. Specific examples include piperidine,piperazine, morpholine, thiamorpholine, thiamorpholine sulfoxide andthiamorpholinesulphone.

[0053] “Heteroalkyl” means an alkyl, cycloalkyl, or cycloalkylalkylradical as defined above, carrying a substituent selected from—NR^(a)R^(b), —OR^(c), or S(O)_(n)R^(d), wherein:

[0054] n is an integer from 0 to 2,

[0055] R^(a) is hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, aralkenyl, heteroaryl, heteroaralkyl, heteroaralkenyl, oracyl;

[0056] R^(b) is hydrogen, alkyl, aryl, aralkyl, acyl, —SO₂R (where R isalkyl, haloalkyl, amino, monosubstituted amino or disubstituted amino),—COOR (where R is alkyl, aralkyl, or heteroaralkyl), —CONR′R″,-(alkylene)CONR′R″ (where R′ and R″ are independently selected fromhydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryland heteroaralkyl, or R′ and R″ together with the nitrogen they areattached to form a cycloamino ring);

[0057] or R^(a) and R^(b) together with the nitrogen atom which they areattached to form a cycloamino ring.

[0058] R^(c) is hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, aralkenyl, heteroaryl, heteroaralkyl, heteroaralkenyl,acyl, —CONR′R″ (where R′ and R″ are independently selected fromhydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryland heteroaralkyl, or R′ and R″ together with the nitrogen they areattached to form a cycloamino ring).

[0059] R^(d) is hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, aralkenyl, heteroaryl, heteroaralkyl, heteroaralkenyl,acyl, -and additionally when n=0, CONR′R″ (where R′ and R″ areindependently selected from hydrogen, alkyl, cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heteroaryl and heteroaralkyl, or R′ andR″ together with the nitrogen they are attached to form a cycloaminoring) and when n=2, NR′R″ where R′ and R″ have the meanings givenimmediately above.

[0060] Representative examples of heteroalkyl include, but are notlimited to 2-methoxyethyl, benzyloxymethyl, thiophen-2-ylthiomethyl, andthe like;

[0061] “Cycloalkylalkyl” means a radical —R^(a)R^(b) where R^(a) is analkylene group and R^(b) is a cycloalkyl group as defined above e.g.,cyclopropylmethyl, cyclohexylpropyl, 3-cyclohexyl-2-methylpropyl, andthe like. “Aralkyl” means a radical —R^(a)R^(b) where R^(a) is analkylene group and R^(b) is an aryl group as defined above e.g., benzyl,phenylethyl, 3-(3-chlorophenyl)-2-methylpentyl, and the like.

[0062] “Aralkenyl” means a radical —R^(a)R^(b) where R^(a) is an alkenylgroup and R^(b) is an aryl group as defined above e.g.,3-phenyl-2-propenyl, and the like.

[0063] “Heteroaralkyl” means a radical —R^(a)R^(b) where R^(a) is analkylene group and R^(b) is a heteroaryl group as defined above e.g.,pyridin-3-ylmethyl, 3-(benzofuran-2-yl)propyl, and the like.

[0064] “Heteroaralkenyl” means a radical —R^(a)R^(b) where R^(a) is analkenyl group and R is a heteroaryl group as defined above e.g.,3-pyridin-3-ylpropen-2-yl, and the like.

[0065] “Heterocycloalkyl” means a radical —R^(a)R^(b) where R^(a) is analkylene group and R^(b) is a heterocyclo group as defined above e.g.,tetrahydropyran-2-ylmethyl, 4-methylpiperazin-1-ylethyl, and the like.

[0066] “Alkoxy”, “aryloxy”, “heteroaryloxy”, “aralkyloxy”, or“heteroaralkyloxy” means a radical —OR where R is an alkyl, aryl,heteroaryl, aralkyl, or heteroaralkyl respectively, as defined abovee.g., methoxy, phenoxy, pyridin-2-yloxy, benzyloxy, and the like.

[0067] “Optional” or “optionally” means that the subsequently describedevent or circumstance may but need not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not. For example, “heterocyclo group optionally mono-or di-substituted with an alkyl group” means that the alkyl may but neednot be present, and the description includes situations where theheterocyclo group is mono- or disubstituted with an alkyl group andsituations where the heterocyclo group is not substituted with the alkylgroup.

[0068] The term “protecting group” refers to a grouping of atoms thatwhen attached to a reactive group in a molecule masks, reduces orprevents that reactivity. Examples of protecting groups can be found inT. W. Greene and P. G. Futs, Protective Groups in Organic Chemistry,(Wiley, 2nd ed. 1991) and Harrison and Harrison et al., Compendium ofSynthetic Organic Methods, Vols. 1-8 (John Wiley and Sons. 1971-1996).Representative amino protecting groups include formyl, acetyl,trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl(Boc), trimethyl silyl (TMS), 2-trimethylsilyl-ethanesulfonyl (SES),trityl and substituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (FMOC), nitro-veratryloxycarbonyl (NVOC)and the like. Representative hydroxy protecting groups include thosewhere the hydroxy group is either acylated or alkylated such as benzyland trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers,trialkylsilyl ethers and allyl ethers.

[0069] The term “protected hydroxylamine derivative” refers to amodified hydroxylamine whose nitrogen and/or hydroxyl groups areprotected such that the nitrogen atom may be selectively monoacylated.

[0070] Compounds that have the same molecular formula but differ in thenature or sequence of bonding of their atoms or the arrangement of theiratoms in space are termed “isomers”. Isomers that differ in thearrangement of their atoms in space are termed “stereoisomers”.Stereoisomers that are not mirror images of one another are termed“diastereomers” and those that are non-superimposable mirror images ofeach other are termed “enantiomers”. When a compound has an asymmetriccenter, for example, it is bonded to four different groups, a pair ofenantiomers is possible. An enantiomer can be characterized by theabsolute configuration of its asymmetric center and is described by theR- and S-sequencing rules of Cahn and Prelog, or by the manner in whichthe molecule rotates the plane of polarized light and designated asdextrorotatory or levorotatory (i.e., as (+) or (−)-isomersrespectively). A chiral compound can exist as either individualenantiomer or as a mixture thereof. A mixture containing equalproportions of the enantiomers is called a “racemic mixture”.

[0071] The compounds of this invention may possess one or moreasymmetric centers; such compounds can therefore be produced asindividual (R)- or (S)-stereoisomers or as mixtures thereof. Forexample, if the R¹ and R² substituents in a compound of formula (I) aredifferent, then the carbon to which they are attached is an asymmetriccenter and therefore the compound of formula (I) can exist as an (R)- or(S)-stereoisomer. Unless indicated otherwise, the description or namingof a particular compound in the specification and claims is intended toinclude both individual enantiomers and mixtures, racemic or otherwise,thereof. The methods for the determination of stereochemistry and theseparation of stereoisomers are well-known in the art (see discussion inChapter 4 of “Advanced Organic Chemistry”, 4th edition J. March, JohnWiley and Sons, New York, 1992).

[0072] A “pharmaceutically acceptable excipient” means an excipient thatis useful in preparing a pharmaceutical composition that is generallysafe, non-toxic and neither biologically nor otherwise undesirable, andincludes an excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

[0073] A “pharmaceutically acceptable salt” of a compound means a saltthat is pharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include:

[0074] (1) acid addition salts, formed with inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like; or formed with organic acids such asacetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid,glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid,malic acid, maleic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelicacid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonicacid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-napthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,4,4′-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionicacid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuricacid, gluconic acid, glutamic acid, hydroxynapthoic acid, salicylicacid, stearic acid, muconic acid, and the like; or

[0075] (2) salts formed when an acidic proton present in the parentcompound either is replaced by a metal ion, e.g., an alkali metal ion,an alkaline earth ion, or an aluminum ion; or coordinates with anorganic base such as ethanolamine, diethanolamine, triethanolamine,tromethamine,-N-methylglucamine, and the like.

[0076] “Pro-drugs” means any compound which releases an active parentdrug according to Formula (I) in vivo when such prodrug is administeredto a mammalian subject. Prodrugs of a compound of Formula (I) areprepared by modifying functional groups present in the compound ofFormula (I) in such a way that the modifications may be cleaved in vivoto release the parent compound. Prodrugs include compounds of Formula(I) wherein a hydroxy, amino, or sulfhydryl group in a compound ofFormula (I) is bonded to any group that may be cleaved in vivo toregenerate the free hydroxyl, amino, or sulfhydryl group, respectively.Examples of prodrugs include, but are not limited to esters (e.g.,acetate, formate, and benzoate derivatives), carbamates (e.g.,N,N-dimethylaminocarbonyl) of hydroxy functional groups in compounds ofFormula (I), and the like.

[0077] “Treating” or “treatment” of a disease includes:

[0078] (1) preventing the disease, i.e. causing the clinical symptoms ofthe disease not to develop in a mammal that may be exposed to orpredisposed to the disease but does not yet experience or displaysymptoms of the disease,

[0079] (2) inhibiting the disease, i.e., arresting or reducing thedevelopment of the disease or its clinical symptoms, or

[0080] (3) relieving the disease, i.e., causing regression of thedisease or its clinical symptoms.

[0081] A “therapeutically effective amount” means the amount of acompound that, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on the compound,the disease and its severity and the age, weight, etc., of the mammal tobe treated.

Nomenclature

[0082] Compounds of this invention may be conveniently named withreference to their amino acid components in accordance with nomenclatureconventional to the peptide field.

[0083] For example a dipeptide where n is 0, R¹=R³=R⁴=R⁶=H, R² is4-thiazolylmethyl, R⁵ is (S,S)-1-methylpropyl and ZR⁷ isbenzyloxycarbonyl, is named as CBz-Ile-4-Taz-NHOH. With respect to thesynthetic schemes A and B subsequently presented, 4-Taz(4-thiazolylalanine) represents AA₁ and Ile represents AA₂.

[0084] A tripeptide where n is 1 and m is 0, R¹=R³=R⁴=R⁶=R⁹=H, R² is4-thiazolylmethyl, R⁵ is (S,S)-1-methylpropyl, ZR⁷ is 4-chlorobenzoyland R¹⁰ is 4-fluorobenzyl is named4-chlorobenzoyl-(4-fluorobenzyl)Gly-Ile-Taz-NHOH. With respect to thesynthetic schemes A and B subsequently presented, 4-Taz represents AA₁,Ile represents AA₂ and (4-fluorobenzyl)Gly represents AA₃.

Preferred Embodiments

[0085] While the broadest definition of this invention is set forth inthe Summary of the Invention, certain compounds of formula (I) arepreferred.

[0086] One class of compounds are the dipeptidic hydroxamic acidscorresponding to compounds of Formula 1 where n is 0. Another class ofcompounds are the tripeptidic hydroxamic acids corresponding tocompounds of Formula 1 where n is 1. For both the dipeptidic andtripeptidic hydroxamic acid compounds of the invention, particularlypreferred are those where R² is heteroaralkyl, particularly4-thiazolylmethyl and R⁵ is (S,S)-1-methylpropyl. Preferably, R², R⁵ andR⁹ are present in the naturally ocurring amino acid configuration, i.e.derived from the (L) amino acids.

[0087] Another class of preferred compounds of Formula I is that where Zis C(O)O. Another class of preferred compounds of Formula I is thatwhere Z is S(O)₂. Within each of these classes of compounds, morepreferred are those where R⁷ is aryl, aralkyl or heteroaryl,particularly halophenyl (e.g. 3,4-dibromophenyl, 2,5-dichlorophenyl or2,4,5-trichlorophenyl), benzyl (optionally substituted) (e.g. benzyl or3,4-dichlorobenzyl) or halothienyl (e.g. 4,5-dibromothien-2-yl). When Zis C═O, also preferred are R⁷ being aralkyl or heteroaralkyl,particularly benzyl or heteroarylmethyl substituted with halo,preferably one or two chloro.

[0088] Another class of preferred compounds is that where R² is aralkylor heteroaralkyl. Within this class, more preferred are those compoundswhere R² is 3-indolyl methyl, 2-thienylmethyl, 4-imidazolylmethyl or4-thiazolylmethyl, particularly 4-thiazolylmethyl.

[0089] Another class of preferred compounds are those where R² is(alkylene)-B—X where B is —O—, —NR⁸—, —S—, —C═O, —CONR⁸—, —NR⁸CO₂—,—NRSO₂— or —C(═NR⁸)NR⁸SO₂— (where R⁸ is independently H or alkyl), and Xis cycloalkyl, cycloalkylalkyl, aryl, aralkyl heteroaryl orheteroaralkyl. Particularly preferred are compounds where the alkylenegroup is methylene, B is —NR⁸CO₂ and X is aralkyl.

[0090] A fourth class of preferred compounds are those where R⁵ is alkylor phenyl, particularly propyl, 1-methylethyl, or (S,S)-1-methylpropyl.Within this class of compounds, particularly preferred are those whereR¹ and R⁴ are hydrogen and R² is heteroaralkyl, particularly4-thiazolylmethyl.

[0091] Representative Compounds of This Invention That Have BeenPrepared According to the Schemes and Examples are as Follows:

[0092] I. Dipeptidic hydroxamic acids of Formula (I) where n=0,R¹=R³=R⁴=R⁶=hydrogen and ZR⁷ is benzyloxycarbonyl and other groups areas defined below are: TABLE I Compound Example St* R2 St** R5 m/e  1 I S4-thiazolylmethyl S phenyl 455  2 I S 4-thiazolylmethyl S 2-carboxyethyl451  3 I S 4-thiazolylmethyl S isobutyl 435  4 I S 4-thiazolylmethyl Sn-butyl 435  5 I S 4-thiazolylmethyl S ethyl 407  6 I S4-thiazolylmethyl S 2-phenethyl 483  7 I S 4-thiazolylmethyl S3-indolylmethyl 508  8 I S 4-thiazolylmethyl S hydroxymethyl 409  9 I S4-thiazolylmethyl S,R 1-benzyloxyethyl 513 10 I S 4-thiazolylmethyl S4-fluorobenzyl 487 11 I S 4-thiazolyimethyl S benzyl 469 12 I S4-thiazolylmethyl S methyl 393 13 I S 4-thiazolylmethyl S1-naphthylmethyl 519 14 I S (E)-styrylmethyl S,S 1-methylpropyl 454 15 IS 2-bromobenzyl S,S 1-methylpropyl 506 16 I S 3-thienylmethyl S,S1-methylpropyl 434 17 I S 5-(2-bromothienyl)- S,S 1-methylpropyl 511methyl 18 I S 2-furylmethyl S,S 1-methylpropyl 418 19 I S3-benzothienylmethyl S,S 1-methylpropyl 484 20 I S 4-fluorobenzyl S,S1-methylpropyl 446 21 I S benzyloxymethyl S,S 1-methylpropyl 458 22 IS,R 1-benzyloxyethyl S,S 1-methylpropyl 472 23 I S 3-chlorobenzyl S,S1-methylpropyl 462 24 I S 4-thiazolylmethyl S cyclopropylmethyl 433 25 IS 4-thiazolylmethyl S cyclohexyl 461 26 I S 4-thiazolylmethyl S2-methylthioethyl 453 27 I S 4-thiazolylmethyl S carbamoylmethyl 436 28I R 3-indolylmethyl S 2-methylthioethyl 485 29 I R 3-indolylmethyl Sbutyl 467 30 I R 3-indolylmethyl S phenyl 487 31 I S 3-indolylmethyl S,S1-methylpropyl 466 32 I S 3-indolylmethyl S 1-naphthylmethyl 551 33 I S3-indolylmethyl R 2-naphthylmethyl 551 34 I S 3-indolylmethyl R2-naphthylmethyl 551 35 I S 3-indolylmethyl S 2-naphthylmethyl 551 36 IS 3-indolylmethyl S 2-phenethyl 515 37 I S 3-indolylmethyl S4-fluorobenzyl 519 38 I S 3-indolylmethyl S ethyl 439 39 I S3-indolylmethyl S isopropyl 453 40 I S 3-indolylmethyl R 2-carboxyethyl483 41 I R 3-indolylmethyl R 2-(N-hydroxy- 498 carbamoyl)ethyl 42 I S(1-benzyl-1H- S,S 1-methylpropyl 508 imidazol-4-yl)methyl 43 I S4-(2,6-dichloro- S,S 1-methylpropyl 602 benzyloxy)benzyl 44 I S2-cyclohexyloxy- S,S 1-methylpropyl 492 carbonylethyl 45 I S 2-phenethylS,S 1-methylpropyl 442 46 I S 4-methylbenzyl- S,S 1-methylpropyl 488thiomethyl 47 I S 2-naphthylmethyl S,S 1-methylpropyl 478 48 I S4-benzyloxy- S,S 1-methylpropyl 543 carbonylaminobutyl 49 I S4-methoxybenzyl S,S 1-methylpropyl 458 50 I S 4-benzyloxybenzyl S,S1-methylpropyl 534 51 I S 4-methoxybenzyl- S,S 1-methylpropyl 504thiomethyl 52 I S 2-(4-methylbenzyl)- S,S 1-methylpropyl 502 thioethyl”53 I R 4-fluorobenzyl S,S 1-methylpropyl 446 54 I R (1-benzyl-1H- S,S1-methylpropyl 509 imidazol-4-yl)methyl 55 I S 4-(2-chlorobenzyl- S,S1-methylpropyl 577 carboxamido)butyl 56 I S (3-benzyloxymethyl- S,S1-methylpropyl 539 3H-imidazol-4-yl) methyl 57 I S (1H-imidazol-4-yl)-S,S 1-methylpropyl 419 methyl 58 I S 2-pyridylmethyl S,S 1-methylpropyl429 59 I S 3-chlorobenzyl S,S 1-methylpropyl 462 60 I S3,4-dichlorobenzyl S,S 1-methylpropyl 496 61 I S 4-nitrobenzyl S,S1-methylpropyl 473 62 I S 4-bromobenzyl S,S 1-methylpropyl 506 63 I S3-trifluoromethyl- S,S 1-methylpropyl 496 benzyl 64 I S 4-phenylbenzylS,S 1-methylpropyl 504 65 I S indan-2-yl S,S 1-methylpropyl 454 66 I Sbenzyloxycarbonyl- S,S 1-methylpropyl 501 aminomethyl 67 I S4-(4-tolylsulfonamido)- S,S 1-methylpropyl 563 butyl 68 I S2-(4-nitrophenyl)ethyl S,S 1-methylpropyl 499 69 I S3-(4-tosylguanidino)- S,S 1-methylpropyl 591 propyl 70 I S4-tertbutylthiobenzyl S,S 1-methylpropyl 516 71 I S 3-tertbutoxybenzylS,S 1-methylpropyl 500 72 I S 4-tertbutoxy- S,S 1-methylpropyl 558carbonylmethylbenzyl 73 I S 4-carbamoylbenzyl S,S 1-methylpropyl 471 74I S 4-acetamidobenzyl S,S 1-methylpropyl 485 75 I S 2-propargyl S,S1-methylpropyl 376 76 I S 2-allyl S,S 1-methylpropyl 378 77 I S4-thiazolylmethyl S,S 1-methylpropyl 435

[0093] II. Dipeptidic hydroxamic acids of Formula (I) where n=0,R¹=R³=R⁴=R⁶=hydrogen and R⁴ is (S,S)-1-methylpropyl and other groups areas defined below are: TABLE II Compound Example St* R2 ZR7 m/e  1 II S4-thiazolylmethyl 4-methoxybenzenesulfonyl 471  2 II S 4-thiazolylmethylbenzenesulfonyl 441  3 II S 4-thiazolylmethyl 2-chlorobenzenesulfonyl474  4 II S 4-thiazolylmethyl 3-chlorobenzenesulfonyl 474  5 II S4-thiazolylmethyl 4-chlorobenzenesulfonyl 474  6 II S 4-thiazolylmethyl2,4-dichlorobenzenesulfonyl 508  7 II S 4-thiazolylmethyl4-fluorobenzenesulfonyl 458  8 II S 4-thiazolylmethyl3-nitrobenzenesulfonyl 485  9 II S 4-thiazolylmethyl4-methylbenzenesulfonyl 454 10 II S 4-thiazolylmethyl 3-trifluoromethyl-508 benzenesulfonyl 11 II S 4-thiazolylmethyl 4-bromo-2,5- 556difluorobenzenesulfonyl 12 II S 4-thiazolylmethyl 5-dimethylamino-1- 533naphthalenesulfonyl 13 II S 4-thiazolylmethyl 4-isopropylbenzenesulfonyl483 14 II S 4-thiazolylmethyl 2,4,6- 483 trimethylbenzenesulfonyl 15 IIS 4-thiazolylmethyl 4-methoxy-2,3,6- 512 trimethylbenzenesulfonyl 16 IIS 4-thiazolylmethyl 2,3,4,5,6- 511 pentamethylbenzenesulfonyl 17 II S4-thiazolylmethyl 4,5-dibromothiophene-2- 604 sulfonyl 18 II S4-thiazolylmethyl 3,4-dibromobenzenesulfonyl 599 19 II S4-thiazolylmethyl 4-chloro-2,5-dimethyl- 503 benzenesulfonyl 20 II S4-thiazolylmethyl 2,4,5-trichlorobenzene- 542 sulfonyl 21 II S4-thiazolytmethyl 4-bromo-2-(trifluoromethoxy)- 604 benzenesulfonyl 22II S 4-thiazolylmethyl 4-methoxynaphthalene-1- 521 sulfonyl 23 II S4-thiazolylmethyl 4-benzenesulfonyl- 586 thiophene2-sulfonyl 24 II S4-thiazolylmethyl 2-chloro-4- 543 (trifluoromethyl)- benzenesulfonyl 25II S 4-thiazolylmethyl 5-chloronaphthalene-1- 525 sulfonyl 26 II S4-thiazolylmethyl 2,2,5,7,8- 567 pentamethylchroman-6- sulfonyl 27 II S4-thiazolylmethyl 4-methoxy-2- 516 nitrobenzenesulfonyl 28 II S4-thiazolylmethyl 1-naphthalenesulfonyl 491 29 II S 4-thiazolylmethyl5-methoxybenzofuran-2- 511 sulfonyl) 30 II S 4-thiazolylmethyl4-tert-amylbenzenesulfonyl 511 31 II S 4-thiazolylmethyl4-(4-chlorophenoxy)- 566 benzenesulfonyl 32 II S 4-thiazolylmethyl2-(pyrid-2-yl)thiophene-5- 523 sulfonyl 33 II S 4-thiazolylmethyl2-{3-[1-methyl-5- 594 (trifluoromethyl)pyrazolyl]}- thiophene-5-sulfonyl34 II S 4-thiazolylmethyl 3,5-dimethylisoxazole-4- 460 sulfonyl 35 II S4-thiazolylmethyl benzofurazan-4-sulfonyl 482 36 III S 3-indolylmethylbenzylcarbamoyl 466 37 IV S 3-indolylmethyl phenylcarbamoyl 452 38 III S3-indolylmethyl benzoyl 437 39 IV S 3-indolylmethyl2-fluorophenylcarbamoyl 470 40 III S 3-indolylmethyl2-methoxyphenylcarbamoyl 482 41 III S 3-indolylmethylcyclohexylcarbamoyl 458 42 III S 3-indolylmethyl 2-phenethylcarbamoyl480 43 III S 3-indolylmethyl 3,5-dichloro- 520 phenylcarbamoyl 44 IV S3-indolylmethyl 2,4-dichlorobenzoyl 505 45 IV S 3-indolylmethyl4-trifluoromethylbenzoyl 505 46 IV S 3-indolylmethyl 3-methylbenzoyl 45147 IV S 3-indolylmethyl cyclohexanoyl 443 48 IV S 4-thiazolylmethyl3-furanoyl 395 49 IV S 4-thiazolylmethyl cyclopentanoyl 397 50 IV S4-thiazolylmethyl 4-aminobenzoyl 420 51 IV S 4-thiazolylmethyl4-hydroxybenzoyl 420 52 IV S 4-thiazolylmethyl 4-dimethylaminobenzoyl448 53 V S 4-thiazolylmethyl 3,4-dichlorobenzyl- 503 oxycarbonyl 54 V S4-thiazolylmethyl 3-chlorobenzyloxycarbonyl 468 55 V S 4-thiazolylmethyl3-nitrobenzyloxycarbonyl 480 56 II S 4-thiazolylmethyl2-(1-naphthylethane)- 519 sulfonyl 57 IV S 4-thiazolylmethyl (R/S)-2-(4-483 chlorophenoxy)propionyl 58 II S 4-thiazolylmethyl2-phenylbenzimidazole-5- 557 sulfonyl 59 IV S 4-thiazolylmethyl3-isoquinolinecarbonyl 456 60 IV S 4-thiazolylmethyl (P/Σ)-α-hydroxy-3-481 nitrophenylacetyl 61 X S 4-thiazolylmethyl 1-carbonylmethyl- 4942,3,4,6,7,8,9,10-octahydro- 1H-pyrimidino[1,2-a]azepin- 5-ylium bromide62 IV S 4-thiazolylmethyl 4-fluorophenoxyacetyl 452 63 II S4-thiazolylmethyl 4-phenylazobenzene- 545 sulfonyl 64 VII S4-thiazolylmethyl 2-(4-chlorophenyl)- 482 ethylcarbamoyl 65 VII S4-thiazolylmethyl 3-phenylpropylcarbamoyl 462 66 X S 4-thiazolylmethyl1-(4-(2-nitrophenyl)- 548 piperazino)methylcarbonyl 67 IX S4-thiazolylmethyl 2-(3-chlorophenyl- 546 sulfonamido)ethylcarbonyl 68 IVS 4-thiazolylmethyl 4-vinylbenzoyl 431 69 IV S 4-thiazolylmethyl2-ethylsulfanylnicotinoyl 466 70 IV S 4-thiazolylmethyl(R/S)-2-hydroxy-3- 449 phenylpropionyl 71 IV S 4-thiazolylmethyl1-hydroxycyclopropyl- 385 carbonyl 72 IV S 4-thiazolylmethyl(R/S)-tetrahydrofuran-3- 399 carbonyl 73 IV S 4-thiazolylmethyl(R)-(-)-2-oxo-4- 430 thiazolidinecarbonyl 74 IV S 4-thiazolylmethyl(S)-2-hydroxy-2-(1H- 439 imidazol-4-yl)propionyl 75 IV S4-thiazolylmethyl 5-chloro-2-thio- 444 phenecarbonyl 76 IV S4-thiazolylmethyl (R/S)-1-hydroxy-(4- 465 methoxyphenyl)acetyl 77 IV S4-thiazolylmethyl (2,6-dihydroxypyrimidin-4-yl) 453 acetyl 78 III S4-thiazolylmethyl cyclohexylcarbamoyl 426 79 I S benzyloxycarbonyla-benzyloxycarbonyl 501 minomethyl 80 II S benzyloxycarbonyla-2,5-dichlorobenzenesulfonyl 575 minomethyl 81 II S benzyloxycarbonyla-3,4-dibromobenzene- 664 minomethyl sulfonyl 82 II S benzyloxycarbonyla-4,5-dibromothiophene-2- 670 minomethyl sulfonyl 83 II Sbenzyloxycarbonyla- 2,5-dimethyl-4- 569 minomethyl chlorobenzenesulfonyl84 II S benzoylaminomethyl benzyloxycarbonyl 471 85 II S4-thiazolylmethyl 2,5-dichlorobenzenesulfonyl 508

[0094] III. Dipeptidic hydroxamic acids of Formula (I) where n=0,R¹=R³=R⁶=hydrogen and ZR⁷ is benzyloxycarbonyl and other groups are asdefined below are: TABLE III Compound Example St* R2 R4 & R5 1 I S3-indolylmethyl propylene 451 2 I S 3-indolylmethyl pentylene 479 3 I S3-indolylmethyl dimethyl 439

[0095] IV. Dipeptidic hydroxamic acids of Formula (I) where n=0,R¹=R³=R⁴=hydrogen, R² is (S)3-indolylmethyl and ZR⁷ is benzyloxycarbonyland other groups are as defined below are: TABLE IV Compound Example St*R5 R6 m/e 1 I H methyl 425 2 I S 2-carboxyethyl methyl 497 3 I S methylmethyl 439 4 I S isopropyl methyl 467 5 I S benzyl methyl 515 6 VI H2-methylpropyl 467 7 VI H 4-pyridylmethyl 502

[0096] V. Tripeptidic hydroxamic acids of Formula I where n=1, m=0,R¹=R³=R⁴=R⁶=H, R² is (S)4-thiazolylmethyl, R⁵ is (S,S)-1-methylpropyland other groups are as defined below are: TABLE V Compound Example St*R9 ZR7 R10 m/e  1 VIII S 2-methylthioethyl benzyloxycarbonyl H 566  2VIII S 2-benzyloxycarbonyl- benzyloxycarbonyl H 654 ethyl  3 VIII S3-indolylmethyl benzyloxycarbonyl H 621  4 VIII S 2-benzyloxyoarbonyl-benzyloxycarbonyl H 654 ethyl  5 VIII S 4-fluorobenzyl benzyloxycarbonylH 600  6 VIII S benzyloxymethyl benzyloxycarbonyl H 612  7 VIII S2-methylpropyl benzyloxycarbonyl H 548  8 VIII S 4-hydroxybenzylbenzyloxycarbonyl H 598  9 VIII S benzyl benzyloxycarbonyl H 582 10 IX Sbenzyl 2-chlorobenzyloxy- H 616 carbonyl 11 VIII S 1-naphthylmethylbenzyloxycarbonyl H 632 12 VIII S phenyl benzyloxycarbonyl H 568 13 VIIIS tert-butoxycarbonyl- benzyloxycarbonyl H 606 methyl 14 VIII S,S1-methylpropyl benzyloxycarbonyl H 548 15 VIII S benzylbenzyloxycarbonyl methyl 596 16 X H 4-chlorobenzoyl 4-fluorobenzyl 60417 X H 3-chlorobenzoyl 4-fluorobenzyl 604 18 X H 3-methylbenzoyl4-fluorobenzyl 584 19 X H phenylcarbamoyl 4-fluorobenzyl 585 20 IX Sbenzyl phenylcarbamoyl H 567 21 X H 4-nitrobenzoyl 4-fluorobenzyl 615 22X H 4-trifluoromethyl- 4-fluorobenzyl 638 benzoyl 23 X H2-methoxybenzoyl 4-fluorobenzyl 600 24 X H 4-(N-hydroxy- 4-fluorobenzyl628 carbamimidoyl)benzoyl 25 X H (R/S) 2-(4- H 525 chlorophenyl)-1-hydroxymethylethyl 26 X H furfuryl H 438 27 X H 2-(4-morpholino)ethyl H471 28 X H 2-pyridylmethyl H 449 29 X H 3-(1-imidazolyl)- H 466 propyl30 X H 2,3-dimethoxybenzyl H 508 31 X H 3-nitrobenzyl H 493 32 X H2-(4-chlorophenethyl) H 496 33 IX S 4-fluorobenzyl 4-tertbutylbenzene- H662 sulfonyl 34 IX S 4-fluorobenzyl 3-chlorobenzene- H 640 sulfonyl 35IX S 4-fluorobenzyl 2,4-dichlorobenzene- H 674 sulfonyl 36 IX S4-fluorobenzyl 4-methoxybenzene- H 636 sulfonyl 37 IX S 4-fluorobenzyl4-methylbenzene- H 620 sulfonyl 38 IX S benzyl 3-chlorobenzene- H 622sulfonyl 39 X H 2,3-dichlorobenzene- 4-fluorobenzyl 674 sulfonyl 40 X H4-tertbutylbenzene- 4-fluorobenzyl 662 sulfonyl

[0097] VI. Miscellaneous tripeptidic hydroxamic acid compounds of theinvention where n=1, m=1, R¹=R³=R⁴=R⁶=H, R² is (S)-4-thiazolylmethyl andR⁵ is (S,S)-1-methylpropyl, and other groups are as defined below areinclude: TABLE VI Compound Example R9 ZR7 R10 m/e 1 IX H 3-bromobenzene-H 591 sulfonyl 2 IX H 3-nitrobenzene- H 557 sulfonyl

[0098] Other miscellaneous compounds with R⁵ and R⁶ forming an alkylenechain include CBz-Pro-Trp-NHOH (m/e=451).

General Synthetic Scheme

[0099] Compounds of this invention may be made by several syntheticmethods as described below.

[0100] The starting materials and reagents used in preparing thesecompounds are either available from commercial suppliers such as AldrichChemical Co., (Milwaukee, Wis., USA), Bachem (Torrance, Calif., USA),Emka-Chemie, or Sigma (St. Louis, Miss., USA) or are prepared by methodsknown to those skilled in the art following procedures set forth inreferences such as Fieser and Fieser's Reagents for Organic Synthesis,Volumes 1-15 (John Wiley and Sons, 1991); Rodd's Chemistry of CarbonCompounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers,1989), Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991),March's Advanced Organic Chemistry, (John Wiley and Sons, 4th Edition),and Larock's Comprehensive Organic Transformations (VCH Publishers Inc.,1989). In particular, a variety of natural and unnatural amino acids invarious protected forms are available from specialty chemical supplierssuch as Novabiochem Inc. (La Jolla, Calif.), Advanced Chemtech Inc.(Louisville, Ky.), Synthetech Inc. (Albany, Oreg.), Bachem Inc.(Torrance, Calif.). These schemes are merely illustrative of somemethods by which the compounds of this invention can be synthesized, andvarious modifications to these schemes can be made and will be suggestedto one skilled in the art having referred to this disclosure.

[0101] The starting materials and the intermediates of the reaction maybe isolated and purified if desired using conventional techniques,including but not limited to filtration, distillation, crystallization,chromatography, and the like.

[0102] The compounds of the invention are C-terminal hydroxamicderivatives of natural and unnatural di- and tripeptides furtherfunctionalized at the N-terminus. They may be made by initiallyassembling the peptide precursor followed by deprotection (as necessary)and functionalization of the C- and N-termini. The peptide precursorsare made by methods known to those of skill in the art of peptidesynthesis, including solution phase chemistries and solid phasesynthesis, see Solid Phase Peptide Synthesis: A Practical Approach by E.Atherton and R. C. Sheppard (Oxford University Press, 1989)

[0103] In general, as shown in Schemes A and B, a C-terminal protectedamino acid P₁—AA₁ is coupled to an N-protected amino acid AA₂—P₂ to givea compound P₁—AA₁—AA₂—P₂. Coupling is accomplished by activating theC-terminus of AA₂—P₂ and condensation with the amine of P₁—AA₁.Activating reagents and coupling conditions are well known to one ofskill in the art and include carbodiimide mediated coupling or formationof N-hydroxysuccinimide esters followed by acylation. For synthesis ofthe dipeptidic hydroxamic acids, protecting group removal and N-terminalfunctionalization with R⁷—Z—L (in either order) is effected to give adipeptide precursor that is converted to a hydroxamic compound of theinvention, typically by treatment with hydroxylamine. For synthesis ofthe tripeptidic hydroxamic acids, compound P₁—AA₁—AA₂—P₂ is selectivelyN-deprotected to give P₁—AA₁—AA₂ which is then coupled to an N-protectedamino acid AA₃—P₃. Deprotection, N-terminal functionalization andtreatment with hydroxylamine as shown in Scheme B then gives tripeptidehydroxamic acids of Formula I.

[0104] With reference to the nomenclature in Formula I, AA₁ correspondsto —C(═O)—CR¹R²—NR³—, AA₂ corresponds to —C(═O)—CR⁴R⁵—NR⁶ and AA₃corresponds to —C(═O)—CH(R⁹)—(CH₂)_(m)—NR¹⁰, P₁, P₂ and P₃ representprotecting groups.

[0105] In particular, compounds of this invention may be prepared bysolid phase synthesis. Initially, an N-protected amino acid P₁—AA₁ isattached to a solid phase resin via its C-terminus. Typical solid phaseresins include chloromethylated and hydroxymethylated resins such as the4-hydroxymethyl-phenylacetamidomethyl resin (Pam resin) and the4-benzyloxybenzyl alcohol resin (Wang resin) available from AdvancedChemtech, Louisville, Ky., U.S.A. and the pegylated polystyrene resin,ArgoGel-OH™, resin from Argonaut Technologies Inc. (Belmont, Calif.).Preferred chloromethyl resins include the styrene/divinylbenzene resinsknown as Merrifield resins available from Aldrich Chemical Company,Milawaukee, Wis., U.S.A.

[0106] Amino acid building blocks, AA₂ and AA₃ are then attachedsequentially using iterative coupling and deprotection steps well knownto one of skill in the art. Coupling reactions are done underconventional peptide coupling conditions, typically in an anhydrousinert aprotic polar solvent (e.g. dimethyl formamide, acetonitrile,tetrahydrofuran, dichloromethane etc.) using auxiliary couplingreagents, e.g., carbodiimides such as1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDAC),diisopropylcarbodiimide (DIC), diimidazoles such as carbonyldiimidazole,triazoles such as hydroxybenzotriazole (HOBT) or other carboxylactivating groups such as N-hydroxysuccinimide, in the presence of antertiary organic base such 4-dimethylaminopyridine, N-methylmorpholineor triethylamine.

[0107] The protecting groups employed depend on the group beingprotected and are also known to those of skill in the art.Representative protecting groups may be found in Protective Groups inOrganic Chemistry, J. F. W. McOmie (London, Plenum Press, (1973)) andProtective Groups in Organic Chemistry, T. W. Greene and P. G. Wuts(John Wiley and Sons, (1991)). A favoured N-protecting group is thefluorenylmethoxycarbonyl (FMOC) group.

[0108] After assembly of the entire peptide skeleton, the protectinggroup at the N-terminus is removed. In certain cases the N-terminalprotecting group may correspond to ZR⁷, obviating the need forprotecting group removal. Functionalization at the N-terminus iseffected by subsequent treatment with a compound R⁷—Z—L, where L is aleaving group under nucleophilic displacement conditions. The conditionsused for protecting group removal vary depending on the protectinggroup. Acid sensitive protecting groups such as tert-butoxycarbonyl(t-BOC) are removed with mild acid (e.g trifluoroacetic acid) whereasbase sensitive protecting groups such as 9-fluorenylmethoxycarbonyl(FMOC) are removed with mild organic base. Typical leaving groups Linclude halo, tosylate and mesylate.

[0109] Finally cleavage off the solid phase is accomplished by treatmentwith hydroxylamine to give compounds of Formula I.

[0110] As stated above, both natural and unnatural amino acids may beused in preparing the compounds of this invention. The natural aminoacids and their abbreviations are well known and will not be repeatedhere. Some examples of unnatural amino acids and their abbreviationsinclude, homoserine (hSer), homoserine lactone (hSerlac), homocysteine(Hcy), homoarginine (hArg), homocitrulline (Hci), penicillamine (Pen),Nα-methylarginine (N-MeArg), norleucine (Nle), norvaline (Nval),norisoleucine (Nile), N-methylisoleucine (N-Meae), phenylglycine (PhG),t-butylglycine (Tle), hydroxyproline (Hyp), 3,4-dehydroproline (A-Pro),pyroglutamine (Pyr,Glp), ornithine (Orn), 2,3-diaminopropionic acid(2,3-DAP), 1-aminoisobutyric acid (1-Aib), 2-aminoisobutyric acid(2-Aib), 2-aminobutyric acid (2-Abu), 4-aminobutyric acid (4-Abu),2,4-diaminobutyric acid (A2bu), a-aminosuberic acid (Asu), albizzin(Abz), β-cyclohexylalanine (Cha), 3-(1-naphthyl)alanine (1-Nal),3-(2-naphthyl)alanine (2-Nal), citrulline (Cit). pipecolinic acid (Pip),4-chlorophenylalanine (4-CIPhe), 4-fluorophenylalanine (4-FPhe),sarcosine (Sar), 4-thiazolylalanine (4-Taz), homophenylalanine (Hpa orhPhe), 2-thienylalanine (2-Thi), 3-benzothienylalanine (3-Bal), and1-aminopropanecarboxylic acid (1-NCPC). A variety of unnatural aminoacids are available from commercial vendors.

Utility Testing, and Administration Utility

[0111] Compounds of this invention are useful to treat diseasesassociated with the excessive deposition of interstitial collagens, suchas fibroproliferatives diseases exemplified by interstitial pulmonaryfibrosis, pericentral fibrosis, Symmers' fibrosis, perimuscularfibrosis, kidney fibrosis, endocardial sclerosis, hepatitis, acuterespiratory distress syndrome, arthritis, cystic fibrosis, surgicaladhesions, tendon surgery, corneal scarring, scleroderma, chronicallograft rejection, hemodialysis shunt fibrosis and restenosis.

[0112] Compounds of this invention are inhibitors of procollagenC-proteinase and thereby inhibit the C-terminal processing of types I,II and III collagens necessary for their ability to form insolublecollagen fibrils. Furthermore, selected compounds of the inventionselectively inhibit procollagen C-proteinase over other collagendegrading enzymes such as collagenase-1, collagenase-2 andcollagenase-3. As a result, compounds of this invention leave largelyunaffected the natural resorption of collagen mediated by collagenase-1,collagenase-2 and collagenase-3. Due to this selectivity, such compoundsare of greater therapeutic efficacy than nonselective inhibitors. Inparticular, preferred compounds of this invention inhibit procollagenC-proteinase with greater than 100 fold selectivity over collagenase-1and collagenase-2 and the most selective compounds are more than athousand fold more selective. Selective inhibition of procollagenC-proteinase over collagenase-1 and collagenase-2 was demonstrated bythe assays described in the Examples. Thereby, this invention allows thetreatment of fibrotic diseases by administering to a patient an agentthat selectively inhibits procollagen C-proteinase over collagenase-1,collagenase-2 and collagenase-3. The inhibition may be 10 fold moreselective, preferably 100 fold more selective and most preferably 1000fold more selective.

Testing

[0113] The ability of the compounds of Formula (I) to inhibitprocollagen C-proteinase activity, may be demonstrated by a variety ofin vitro assays known to those of ordinary skill in the art, such as theassay described in more detail in Example XIII. The selectivity againstcollagenase enzymes may be determined by testing as described in ExampleXIV.

[0114] The in vivo efficacy of compounds of Formula (I) against fibroticdisease and the overproduction and deposition of collagen may be shownby numerous animal models including the mouse bleomycin inducedpulmonary fibrosis model (S. H. Phan et.al. “Bleomycin-induced PulmonaryFibrosis,” Am. Rev. Respir. Dis., 124:428-434 (1981) and P. F. Piguet etal. “Effective Treatment of the Pulmonary Fibrosis Elicited in Mice byBleomycin or Silica with anti-CD-11 Antibodies,” Am. Rev. Resp. Dis.,147:435-441 (1993)) the sponge implant model (E. N. Unemori et al.“Human Relaxin Decreases Collagen Accumulation In Vivo in Two RodentModels of Fibrosis,” J. Invest. Dermatol., 101:280-285 (1993), thecarbon tetrachloride or NDMU induced renal fibrosis model, as well asother animal models cited in WO 97/05865 (“C-Proteinase Inhibitors forthe Treatment of Disorders Relating to the Overproduction of Collagen”),published Feb. 20, 1997.

Administration and Pharmaceutical Composition

[0115] In general, the compounds of this invention will be administeredin a therapeutically effective amount by any of the accepted modes ofadministration for agents that serve similar utilities. The actualamount of the compound of this invention, i.e., the active ingredient,will depend upon numerous factors such as the severity of the disease tobe treated, the age and relative health of the subject, the potency ofthe compound used, the route and form of administration, and otherfactors. The drug can be administered more than once a day, preferablyonce or twice a day.

[0116] Therapeutically effective amounts of compounds of formula I mayrange from approximately 0.05-35 mg per kilogram body weight of therecipient per day; preferably about 0.3-20 mg/kg/day. Thus, foradministration to a 70 kg person, the dosage range would most preferablybe about 21 mg to 1.4 g per day.

[0117] In general, compounds of this invention will be administered aspharmaceutical compositions by any one of the following routes: oral,systemic (e.g., transdermal, intranasal, pulmonary or by suppository),or parenteral (e.g., intramuscular, intravenous or subcutaneous)administration. The preferred manner of administration is systemic usinga convenient daily dosage regimen which can be adjusted according to thedegree of affliction.

[0118] Intranasal delivery is typically accomplished with dry powderformulations, liquid solutions or suspensions suitable for nebulizationor with aerosol propellants suitable for use in a metered dose inhaler.Alternatively, drug substance may be associated with microspheres madeof materials such as gelatin, dextran, collagen or albumin Themicrospheres are conveniently delivered in freeze dried form with anasal insufflator device or a pressurized aerosol cannister.

[0119] Penetration enhancers such as amphiphilic steroids may also beused as additives to increase the systemic absorption of the drug intothe tissue. Effective administration may also be accomplished bypulmonary or respiratory delivery since polypeptides are readilyabsorbed through the cellular lining of the alveolar region of themammalian lung. Advantageously, such administration frequently does notrequire the use of penetration enhancers as additives. Devices andmethods for pulmonary delivery deep into the lung are described in U.S.Pat. No. 5,780,014, issued Jul. 14, 1998 and U.S. Pat. No. 5,814,607,issued Sep. 29, 1998.

[0120] Lastly, compounds may be systemically administered by transdermaldelivery, which typically involves placing the drug on the surface ofthe skin and allowing it to permeate through the skin. Transdermaldelivery devices employ a structure such as an adhesive patch or thelike that serves as a reservoir for the drug and brings the drug intodiffusive contact with the skin. In one general typ, the structure is athree dimensionally stable matrix known as a monolithic matrix. Suchmatrices are described in more detail in U.S. Pat. Nos. 5,804,214,5,149,538 and 4,956,171 which describe matrices made of polymers andcopolymers of acrylic latexes, acrylic esters, methacrylic esters andvinyl acetates.

[0121] The choice of formulation depends on various factors such as themode of drug administration (e.g., for oral administration, formulationsin the form of tablets, pills or capsules are preferred) and thebioavailability of the drug substance. Recently, pharmaceuticalformulations have been developed especially for drugs that show poorbioavailability based upon the principle that bioavailability can beincreased by increasing the surface area i.e., decreasing particle size.For example, U.S. Pat. No. 4,107,288 describes a pharmaceuticalformulation having particles in the size range from 10 to 1,000 nm inwhich the active material is supported on a crosslinked matrix ofmacromolecules. U.S. Pat. No. 5,145,684 describes the production of apharmaceutical formulation in which the drug substance is pulverized tonanoparticles (average particle size of 400 nm) in the presence of asurface modifier and then dispersed in a liquid medium to give apharmaceutical formulation that exhibits remarkably highbioavailability.

[0122] The compositions are comprised of in general, a compound offormula (I) in combination with at least one pharmaceutically acceptableexcipient. Acceptable excipients are non-toxic, aide administration, anddo not adversely affect the therapeutic benefit of the compound offormula (I). Such excipient may be any solid, liquid, semi-solid or, inthe case of an aerosol composition, gaseous excipient that is generallyavailable to one of skill in the art.

[0123] Solid pharmaceutical excipients include starch, cellulose, talc,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, magnesium stearate, sodium stearate, glycerol monostearate, sodiumchloride, dried skim milk and the like. Liquid and semisolid excipientsmay be selected from glycerol, propylene glycol, water, ethanol andvarious oils, including those of petroleum, animal, vegetable orsynthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesameoil, etc. Preferred liquid carriers, particularly for injectablesolutions, include water, saline, aqueous dextrose, and glycols.

[0124] Compressed gases may be used to disperse a compound of thisinvention in aerosol form. Inert gases suitable for this purpose arenitrogen, carbon dioxide, etc.

[0125] Other suitable pharmaceutical excipients and their formulationsare described in Remington's Pharmaceutical Sciences, edited by E. W.Martin (Mack Publishing Company, 18th ed., 1990).

[0126] The amount of the compound in a formulation can vary within thefull range employed by those skilled in the art. Typically, theformulation will contain, on a weight percent (wt %) basis, from about0.01-99.99 wt % of a compound of formula I based on the totalformulation, with the balance being one or more suitable pharmaceuticalexcipients. Preferably, the compound is present at a level of about 1-80wt %. Representative pharmaceutical formulations containing a compoundof formula I are described in Example 30.

EXAMPLES

[0127] The following preparations and examples are given to enable thoseskilled in the art to more clearly understand and to practice thepresent invention. They should not be considered as limiting the scopeof the invention, but merely as being illustrative and representativethereof. Ac₂Ol capping solution—19 mL Ac₂O, 9 mL DIPEA, 0.8 eq. HOBt,400 mL NMP

[0128] Abbreviations

[0129] Ac₂O—acetic anhydride

[0130] DIPEA—diisopropylethylamine

[0131] HOBt—hydroxybenzotriazole

[0132] NMP—N-Methylpyrrolidinone

[0133] FMOC—Fluorenylmethoxycarbonyl

[0134] BOC—t-butoxycarbonyl

[0135] DIC—diisopropylcarbodiimide

[0136] DMAP—4-dimethylaminopyridine

[0137] HATU—O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate]

[0138] HOAT—1-hydroxy-7-azabenzotriazole

[0139] TFA—trifluoroacetic acid

[0140] DMF—dimethylformamide

[0141] TIS—triisopropylsilane

[0142] CBz—benzyloxycarbonyl

[0143] THF—tetrahydrofuran

[0144] Taz—4-thiazolylalanine

[0145] Ile—isoleucine

[0146] Su—succinimidyl

[0147] TEA—triethylamine

[0148] Trp—tryptophan

[0149] DMSO—dimethylsulfoxide

[0150] TMS—trimethylsilyl

[0151] DMEM-HG—Dulbeccos Modified Eagle Medium, high glucose

[0152] G418—Geniticin

Synthesis of Compounds of Formula I

[0153] Compounds of Formula (I) were made by solid phase synthesis usingconventional methods as generally described in the following Examples.If necessary, the compounds prepared as described in Examples I-X werepurified by reverse phase high pressure liquid chromatography on silicagel bonded diisopropylphenethylsilane colunumns (Zorbax SB-Phenyl) usinggradient elution with a mixed acetonitrile-water (1% TFA) solvent systemat flow rates of approximately 1.5-2.0 mL/minute. They were directlytested in the PCP and collagenase assays described in Examples XIII andXIV without further purification. The compounds may be characterizedusing conventional means, including physical constants and spectraldata. In particular, they were analyzed by mass spectrometry usingelectron spray ionization.

Example I General Experimental for CBz—AA₂—AA₁—NHOH (Dipeptide Compoundsof Formula I where ZR⁷ is benzyloxycarbonyl)

[0154] 1. BOC-AA₁ or FMOC-AA₁

[0155] To ArgoGel-OH™ (Argonaut Technologies, Belmont, Calif.) in aempty solid phase extraction vial, fitted with a stopcock was added 3eq. of either BOC or FMOC protected AA₁, 3 eq. ofdiisopropylcarbodiimide and 0.05 eq. of a 0.116 M solution ofdimethylaminopyridine in THF. Sufficient CH₂Cl₂ was added to swell theresin (˜12.5 mL/gr of resin). [Alternatively, to the resin was added 3eq. of either BOC or FMOC protected AA₁, 3 eq. of HATU, 3 eq. HOAt and 6eq. of DIPEA. Sufficient NMP was added to swell the resin (˜12.5 mL/grof resin ).] The reaction was then placed on a spinner and rotatedovernight. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, once with 1:1HOAc/CH₂Cl₂, three times with MeOH and then lastly three times withCH₂Cl₂ and dried to give BOC-AA₁-resin or FMOC-AA₁ resin. The unreactedresin sites were then capped off with an Ac₂O capping solution (19 mLAc₂O, 9 mL DIPEA, 0.8 gr HOBt, 400 mL NMP) in a sufficient quantityadded to swell the resin (˜12.5 mL/gr of resin). The resin was thenfiltered and washed as above.

[0156] 2. H₂N—AA₁-resin

[0157] 2a. For BOC Protected Material

[0158] The resulting resin from above (BOC-AA₁-resin) was treated with asolution of 95/2.5/2.5 TFA/H₂O/triisopropyl silane (TIS) for two hours.The reaction was then filtered by suction filtration and washed threetimes with CH₂Cl₂, three times with MeOH, and then three times withCH₂Cl₂ to give H₂N—AA₁-resin.

[0159] 2b. For FMOC Protected Material

[0160] The resulting resin from above (FMOC-AA₁-resin) was first washedwith DMF then treated with a solution of 20% piperidine in DMF for 20min. The reaction was then filtered by suction filtration and washedthree times with CH₂Cl₂, three times with MeOH, once with 1:1HOAc/CH₂Cl₂, three times with MeOH and then lastly three times withCH₂Cl₂, to give H₂N—AA₁-resin.

[0161] 3. CBz—AA₂—AA₁ Resin

[0162] To H₂N—AA₁-resin was added 3 eq. of CBZ protected AA₂, 3 eq. ofdiisopropylcarbodiimide and 0.05 eq. of a 0.116 M solution ofdimethylaminopyridine in THF. Sufficient CH₂Cl₂ was added to swell theresin (˜12.5 mL/gr of resin). [Alternatively, to the resin was added 3eq. of CBz protected AA₂, 3 eq. of HATU, 3 eq. HOAt and 6 eq. of DIPEA.Sufficient NMP is added to swell the resin (˜12.5 mL/gr of resin ).] Thereaction was then placed on a spinner and rotated overnight. Thereaction was then filtered by suction filtration and washed three timeswith CH₂Cl₂, three times with MeOH, once with 1:1 HOAc/CH₂Cl₂, threetimes with MeOH and then lastly three times with CH₂Cl₂ to giveCBz—AA₂—AA₁-resin.

[0163] 4. Removal of Protecting Groups

[0164] If AA₁ or AA₂ contains an acid labile side chain protecting groupthat is to be removed, the resulting resin (CBz—AA₂—AA₁-resin) wastreated with a solution of 95/2.5/2.5 TFA/H₂O/triisopropyl silane (TIS)for 2 hours. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, and then threetimes with CH₂Cl₂.

[0165] 5. CBz—AA₂—AA₁—NHOH

[0166] CBz—AA₂—AA₁-resin was first washed with THF. Sufficient THF wasadded to swell the resin (˜12.5 mL/gr of resin) then 25 eq. of 50% aq.NH₂OH was added and the reaction was rotated for two days. The reactionwas then filtered by suction filtration and washed with CH₂Cl₂, MeOH,and then CH₂Cl₂. The filtrate was concentrated under vacuum to obtainCBz—AA₂—AA₁—NHOH.

Example II General Experimental for R⁷—SO₂—AA₂—AA₁—NHOH (DipeptideCompounds of Formula I where Z is —SO₂—)

[0167] 1. FMOC-AA₂—AA₁-resin

[0168] To H₂N—AA₁-resin was added 3 eq. of FMOC protected AA₂, 3 eq. ofdiisopropylcarbodimide and 0.05 eq. of a 0.116 M solution of4-dimethylaminopyridine in THF. Sufficient CH₂Cl₂ was added to swell theresin (˜12.5 mL/gr of resin). The reaction was then placed on a spinnerand rotated overnight. The reaction was then filtered by suctionfiltration and washed three times with CH₂Cl₂, three times with MeOH,once with 1:1 HOAc/CH₂Cl₂, three times with MeOH and then lastly threetimes with CH₂Cl₂, to give FMOC-AA₂—AA₁-resin.

[0169] 2. H₂N—AA₂—AA₁-resin

[0170] FMOC-AA₂—AA₁-resin was first washed with DMF then treated with asolution of 20% piperidine in DMF for 20 min. The reaction was thenfiltered by suction filtration and washed three times with CH₂Cl₂, threetimes with MeOH, once with 1:1 HOAc/CH₂Cl₂, 3 times with MeOH and thenlastly 3 times with CH₂Cl₂, to give H₂N—AA₂—AA₁-resin.

[0171] 3. R⁷—SO₂—AA₂—AA₁-resin

[0172] H₂N—AA₂—AA₁-resin was first washed with aq. dioxane, then 10 eq.of the desired sulfonyl chloride, R⁷—SO₂Cl, was added. Sufficient 90%aq. dioxane was added to swell the resin (˜12.5 mL/gr of resin). Then 20eq. of diisopropylethylamine was added. The reaction was then placed ona spinner and rotated overnight. The reaction was then filtered bysuction filtration and washed three times with CH₂Cl₂, three times withMeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOH and then lastlythree times with CH₂Cl₂ to give R⁷—SO₂—AA₂—AA₁-resin.

[0173] 4. R⁷—SO₂—AA₂—AA₁—NHOH

[0174] R⁷—SO₂—AA₂—AA₁-resin was first washed with THF. Sufficient THFwas added to swell the resin (˜12.5 mL/gr of resin) then 25 eq. of 50%aq. NH₂OH was added and the reaction was rotated for two days. Thereaction was then filtered by suction filtration and washed with CH₂Cl₂,MeOH, and then CH₂Cl₂. The filtrate was concentrated under vacuum toobtain R⁷—SO₂—AA₂—AA₁—NHOH.

Example III General Experimental for R⁷NHCO—AA₂—AA₁—NHOH (DipeptideCompounds of Formula I where Z is —CONH—)

[0175] 1. R⁷—NHCO—AA₂—AA₁-resin

[0176] H₂N—AA₂—AA₁-resin was first washed with THF, then 3 eq. of thedesired isocyanate, R⁷N═C═O, was added. Sufficient THF was added toswell the resin (˜12.5 mL/gr of resin). The reaction was then placed ona spinner and rotated overnight. The reaction was then filtered bysuction filtration and washed three times with CH₂Cl₂, three times withMeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOH and then lastlythree times with CH₂Cl₂ to give R⁷—NHCO—AA₂—AA₁-resin.

[0177] 2. Removal of Protecting Groups

[0178] If AA₁ or AA₂ contains an acid labile side chain protecting groupthat is to be removed, the resulting resin (R⁷—NHCO—AA₂—AA₁-resin) wastreated with a solution of 95/2.5/2.5 TFA/H₂O/triisopropyl silane (TIS)for 2 hours. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, and then threetimes with CH₂Cl₂.

[0179] 3. R⁷—NHCO—AA₂—AA₁—NHOH

[0180] R⁷NHCO—AA₂—AA₁-resin was first washed with THF. Sufficient THFwas added to swell the resin (˜12.5 mL/gr of resin) then 25 eq. of 50%aq. NH₂OH was added and the reaction was rotated for two days. Thereaction was then filtered by suction filtration and washed with CH₂Cl₂,MeOH, and then CH₂Cl₂. The filtrate was concentrated under vacuum toobtain R⁷NHCO—AA₂—AA₁—NHOH.

[0181] The dipeptide hydroxamic acids in Table II and Table V where Z is—CONH— were prepared using the procedure of Example 3.

Example IV General Experimental for R⁷—CO—AA₂—AA₁—NHOH (DipeptideCompounds of Formula I where Z is —CO—)

[0182] 1. R⁷—CO—AA₂—AA₁-resin

[0183] To H₂N—AA₂—AA₁-resin was added sufficient CH₂Cl₂ to swell theresin (˜12.5 mL/gr of resin). 3 eq. of the desired acid chloride,R⁷COCl, and 3 eq. of Et₃N were then added. A second alternative was addto the CH₂Cl₂ swollen resin 3 eq. of the desired carboxylic acid,R⁷CO₂H, 3 eq. of diisopropylcarbodiimide and 0.05 eq. of a 0.116 Msolution of dimethylaminopyridine in THF. A third alternative was toswell the resin with sufficient CH₃CN (˜12.5 mL/gr of resin) and couplethe desired acid chloride, R⁷COCl (3 eq.) in the presence of 6 eq. oftrimethylsilylcyanide. In all cases the reaction was then placed on aspinner and rotated overnight. The reaction was then filtered by suctionfiltration and washed three times with CH₂Cl₂, three times with MeOH,once with 1:1 HOAc/CH₂Cl₂, three times with MeOH and then lastly threetimes with CH₂Cl₂ to give R⁷—CO—AA₂—AA₁-resin.

[0184] 2. Removal of Protecting Groups

[0185] If AA₁ or AA₂ contains an acid labile side chain protecting groupthat is to be removed, the resulting resin (R⁷—CO—AA₂—AA₁-resin) wastreated with a solution of 95/2.5/2.5 TFA/H₂O/triisopropyl silane (TIS)for 2 hours. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, and then threetimes with CH₂Cl₂.

[0186] 3. R⁷—CO—AA₂—AA₁—NHOH

[0187] R⁷—CO—AA₂—AA₁-resin was first washed with THF. Sufficient THF wasadded to swell the resin (˜12.5 mL/gr of resin) then 25 eq. of 50% aq.NH₂OH was added and the reaction was rotated for two days. The reactionwas then filtered by suction filtration and washed with CH₂Cl₂, MeOH,and then CH₂Cl₂. The filtrate is concentrated under vacuum to obtainR⁷—CO—AA₂—AA₁—NHOH.

Example V General Experimental for R⁷—OC(═O)—AA₂—AA₁—NHOH (DipeptideCompounds of Formula I where Z is —C(O)O—)

[0188] 1. R⁷—OCO—AA₂—AA₁-resin.

[0189] To H₂N—AA₂—AA₁-resin was added 3 eq. of the desiredsuccinimidylcarbonate, R⁷OC(═O)NHS, 3 eq. Et₃N and and 0.05 eq. of a0.116 M solution of 4-dimethylaminopyridine in THF. Sufficient CH₂Cl₂was added to swell the resin (˜12.5 mL/gr of resin). Another alternativewas to add to the resin 10 eq. of the desired chloroformate, R⁷OCOCl,and 20 eq. diisopropylethylamine followed by sufficient 90% aq. dioxaneto swell the resin (˜12.5 mL/gr of resin)]. In either case, the reactionwas then placed on a spinner and rotated overnight. The reaction wasthen filtered by suction filtration and washed three times with CH₂Cl₂,three times with MeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOHand lastly three times with CH₂Cl₂ to give R⁷—OCO—AA₂—AA₁-resin.

[0190] 2. Removal of Protecting Groups

[0191] If AA₁ or AA₂ contains an acid labile side chain protecting groupthat is to be removed, the resulting resin (R⁷—OCO—AA₂—AA₁-resin) wastreated with a solution of 95/2.5/2.5 TFA/H₂O/triisopropyl silane (TIS)for 2 hours. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, and then threetimes with CH₂Cl₂.

[0192] 3. R⁷—OCO—AA₂—AA₁—NHOH

[0193] R⁷—OCO—AA₂—AA₁-resin was first washed with THF. Sufficient THFwas added to swell the resin (˜12.5 mL/gr of resin) and 25 eq. of 50%aq. NH₂OH is added and the reaction was rotated for two days. Thereaction was then filtered by suction filtration and washed with CH₂Cl₂,MeOH, and then CH₂Cl₂. The filtrate was concentrated under vacuum toobtain R⁷—OCO—AA₂—AA₁—NHOH.

Example VI General Experimental for CBz—NR⁶—CH₂CO—AA₁—NHOH

[0194] 1. BrCH₂CO—AA₁-resin

[0195] To H₂N—AA₁-resin was added 12 eq. of bromoacetic acid and 13 eq.of diisopropylcarbodiimide. Sufficient CH₂Cl₂ was added to swell theresin (˜12.5 mL/gr of resin). The reaction was then placed on a spinnerand rotated for 2 hrs. The reaction was filtered by suction filtrationand washed three times with CH₂Cl₂, three times with DMSO and lastlythree times with CH₂Cl₂ to give BrCH₂CO—AA₁-resin.

[0196] 2. R⁶—NHCH₂CO—AA₁-resin

[0197] BrCH₂CO—AA₁-resin was first washed with DMSO. Sufficient DMSO wasadded to swell the resin (˜12.5 mL/gr of resin) followed by the additionof 40 eq. of the desired primary amine, R⁶NH₂. The reaction was thenplaced on a spinner and rotated overnight. The reaction was thenfiltered by suction filtration and washed three times with CH₂Cl₂, threetimes with MeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOH andthen lastly three times with CH₂Cl₂ to give R⁶—NHCH₂CO—AA₁-resin.

[0198] 3. CBz—NR⁶—CH₂CO—AA₁-resin

[0199] To R⁶—NHCH₂CO—AA₁-resin is added 3 eq. of CBZOSu, 3 eq. Et₃N and0.05 eq. of a 0.116 M solution of 4-dimethylaminopyridine in THF.Sufficient CH₂Cl₂ was added to swell the resin (˜12.5 mL/gr of resin).The reaction was then placed on a spinner and rotated overnight. Thereaction was then filtered by suction filtration and washed three timeswith CH₂Cl₂, three times with MeOH, once with 1:1 HOAc/CH₂Cl₂, threetimes with MeOH and lastly three times with CH₂Cl₂ to giveCBz—NR⁶—CH₂CO—AA₁-resin.

[0200] 4. Removal of Protecting Groups

[0201] If AA₁ or AA₂ contains an acid labile side chain protecting groupthat is to be removed, the resulting resin (CBz—NR⁶—CH₂CO—AA₁-resin) wastreated with a solution of 95/2.5/2.5 TFA/H₂O/triisopropyl silane (TIS)for 2 hours. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, and then threetimes with CH₂Cl₂.

[0202] 5. CBZ—NR⁶—CH₂CO—AA₁—NHOH

[0203] CBz—NR⁶—CH₂CO—AA₁-resin was first washed with TBF. Sufficient THFwas added to swell the resin (˜12.5 mL/gr of resin) and then 25 eq. of50% aq. NH₂OH was added and the reaction was rotated for two days. Thereaction was then filtered by suction filtration and washed with CH₂Cl₂,MeOH and finally CH₂Cl₂. The filtrate was concentrated under vacuum toobtain CBz—NR⁶—CH₂CO—AA₁—NHOH.

Example VII General Experimental for R⁷NHCO—AA₂—AA₁—NHOH (DipeptideCompounds of Formula I where Z is —C(O)NH—)

[0204] 1. To ArOCO—AA₂—AA₁-resin (Ar=Ph) was added sufficient DMF toswell the resin (˜12.5 mL/gr of resin). 20 eq. of R⁷NH₂ was added andthe reaction reaction was then placed on a spinner and rotatedovernight. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, once with 1:1HOAc/CH₂Cl₂, three times with MeOH and then lastly three times withCH₂Cl₂ to give R⁷NHCO—AA₂—AA₁-resin.

[0205] 2. R⁷NHCO—AA₂—AA₁-resin was first washed with THF. Sufficient THFwas added to swell the resin (˜12.5 mL/gr of resin), then 25 eq. of 50%aq. NH₂OH was added and the reaction was rotated for two days. Thereaction was then filtered by suction filtration and washed with CH₂Cl₂,MeOH, and then CH₂Cl₂. The filtrate was concentrated under to obtainR⁷NHCO—AA₂—AA₁—NHOH.

Example VIII General Experimental for CBz—AA₃—AA₂—AA₁—NHOH (TripeptideCompounds of Formula I where n=1 and ZR⁷ is benzyloxycarbonyl)

[0206] 1. CBz—AA₃—AA₂—AA₁-resin

[0207] To H₂N—AA₂—AA₁-resin was added three eq. of CBZ protected AA₃,three eq. of diisopropylcarbodiimide and 0.05 eq. of a 0.116 M solutionof dimethylaminopyridine in THF. Sufficient CH₂Cl₂ is added to swell theresin (˜12.5 mL/gr of resin). [An alternative is to add to the resinthree eq. of CBz protected AA₃, three eq. of HATU, three eq. HOAt andsix eq. of DIPEA. Sufficient NMP was added to swell the resin (˜12.5mL/gr of resin).] The reaction was placed on a spinner and rotatedovernight. The reaction was filtered by suction filtration and washedthree times with CH₂Cl₂, three times with MeOH, once with 1:1HOAc/CH₂Cl₂, three times with MeOH and lastly three times with CH₂Cl₂,to give CBz—AA₃—AA₂—AA₁-resin.

[0208] 2. Removal of Protecting Groups

[0209] If AA₁, AA₂ or AA₃ contains an acid labile side chain protectinggroup that is to be removed, the resulting resin (CBz—AA₃—AA₂—AA₁-resin)was treated with a solution of 95/2.5/2.5 TFA/H₂O, triisopropyl silane(TIS) for 2 hours. The reaction was then filtered by suction filtrationand washed three times with CH₂Cl₂, three times with MeOH, and thenthree times with CH₂Cl₂.

[0210] 3. CBz—AA₃—AA₂—AA₁—NHOH

[0211] CBz—AA₃—AA₂—AA₁-resin was first washed with THF. Sufficient THFwas added to swell the resin (˜12.5 mL/gr of resin) then 25 eq. of 50%aq. NH₂OH was added and the reaction was rotated for two days. Thereaction was filtered by suction filtration and washed with CH₂Cl₂,MeOH, and finally with CH₂Cl₂. The filtrate was concentrated undervacuum to obtain CBz—AA₃—AA₂—AA₁—NHOH.

Example IX General Experimental for R⁷Z—AA₃—AA₂—AA₁—NHOH (TripeptideCompounds of Formula I where Z is —SO₂— or —C(O)NH—)

[0212] 1. FMOC-AA₃—AA₂—AA₁-resin

[0213] To H₂N-AA₂-AA,-resin was added three eq. of FMOC protected AA₃,three eq. of diisopropylcarbodiimide and 0.05 eq. of a 0.116 M solutionof 4-dimethylaminopyridine in THF. Sufficient CH₂Cl₂ was added to swellthe resin (˜12.5 mL/gr of resin). [and alternative was to add to theresin three eq. of FMOC protected AA₃, three eq. of HATU, three eq. HOAtand six eq. of DIPEA. Sufficient NMP was added to swell the resin (˜12.5mL/gr of resin).] The reaction was placed on a spinner and rotatedovernight. The reaction was then filtered by suction filtration andwashed three times with CH₂Cl₂, three times with MeOH, once with 1:1HOAc/CH₂Cl₂, three times with MeOH and then lastly three times withCH₂Cl₂ to give FMOC-AA₃—AA₂—AA₁-resin.

[0214] 2. H₂N—AA₃—AA₂—AA₁-resin

[0215] FMOC-AA₃—AA₂—AA₁-resin was first washed with DMF then treatedwith a solution of 20% piperidine in DMF for 20 min. The reaction wasthen filtered by suction filtration and washed three times with CH₂Cl₂,three times with MeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOHand then lastly three times with CH₂Cl₂ to give H₂N—AA₃—AA₂—AA₁-resin.

[0216] 3. R⁷Z—AA₃—AA₂—AA₁-resin

[0217] 3a. For Z=SO₂:

[0218] H₂N—AA₃—AA₂—AA₁-resin is first washed with aq. dioxane, then 10eq. of the desired sulfonyl chloride, R⁷SO₂Cl, is added followed bysufficient 90% aq. dioxane to swell the resin (˜12.5 mL/gr of resin).Then 20 eq. of diisopropylethylamine was added. The reaction was thenplaced on a spinner and rotated overnight. The reaction was thenfiltered by suction filtration and washed three times with CH₂Cl₂, threetimes with MeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOH andthen lastly three times with CH₂Cl₂ to give R⁷SO₂—AA₃—AA₂—AA₁-resin.

[0219] 3b. For Z=NHCO:

[0220] H₂N—AA₃—AA₂—AA₁-resin was first washed with THF and then threeeq. of the desired isocyanate, R⁷N═C═O was added followed by sufficientTHF to swell the resin (˜12.5 mL/gr of resin). The reaction was thenplaced on a spinner and rotated overnight. The reaction was thenfiltered by suction filtration and washed three times with CH₂Cl₂, threetimes with MeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOH andthen lastly three times with CH₂Cl₂ to give R⁷NHCO—AA₃—AA₂—AA₁-resin.

[0221] 4. R⁷Z—AA₃—AA₂—AA₁—NHOH

[0222] R⁷Z—AA₃—AA₂—AA₁-resin is first washed with THF. Sufficient THF isadded to swell the resin (˜12.5 mL/gr of resin) then 25 eq. of 50% aq.NH₂OH is added and the reaction is rotated for two days. The reaction isthen filtered by suction filtration and washed with CH₂Cl₂, MeOH, andthen CH₂Cl₂. The filtrate is concentrated on a Speed Vac to obtainR⁷Z—AA₃—AA₂—AA₁—NHOH.

Example X General Experimental for R⁷Z—NR¹⁰C(O)CH₂—AA₂—AA₁—NHOH(Tripeptide Compounds of Formula I where n=1, m=0, R⁹ is hydrogen (A isC(O)CH₂NR¹⁰) and Z is a bond, —SO₂—, —C═O, or —C(O)NH—)

[0223] 1. BrCH₂CO—AA₂—AA₁-resin

[0224] To H₂N—AA₂—AA₁-resin was added twelve eq. of bromoacetic acid andtwelve eq. of diisopropylcarbodimide. Sufficient CH₂Cl₂ was added toswell the resin (˜12.5 mL/gr of resin). The reaction was then placed ona spinner and rotated two hrs. The reaction was then filtered by suctionfiltration and washed three times with CH₂Cl₂, three times with DMSO andthen lastly three times with CH₂Cl₂ to give BrCH₂CO—AA₂—AA₁-resin.

[0225] 2. R¹⁰NHCH₂CO—AA₂—AA₁-resin

[0226] BrCH₂CO—AA₂—AA₁-resin is first washed with DMSO. Sufficient DMSOis added to swell the resin (˜12.5 mL/gr of resin) followed by theaddition of 40 eq. of the desired amine, R¹⁰NH₂ or R⁷R¹⁰NH. The reactionwas then placed on a spinner and rotated overnight. The reaction wasthen filtered by suction filtration and three times with CH₂Cl₂, threetimes with MeOH, once with 1: 1 HOAc/CH₂Cl₂, three times with MeOH andthen lastly three times with CH₂Cl₂ to give R¹⁰NHCH₂CO—AA₂—AA₁-resin orR⁷R¹⁰NHCH₂CO—AA₂—AA₁-resin. (If R⁷R¹⁰NH is used, corresponding to Zbeing a bond, go directly to step 4)

[0227] 3. R⁷Z—NR¹⁰CH₂CO—AA₂—AA₁-resin

[0228] 3a. For Z=—SO₂—:

[0229] R¹⁰NHCH₂CO—AA₂—AA₁-resin is first washed with aq. dioxane, then10 eq. of the desired sulfonyl chloride, R⁷SO₂Cl, is added. Sufficient90% aq. dioxane was added to swell the resin (˜12.5 mL/gr of resin).Then 20 eq. of diisopropylethylamine was added. The reaction was thenplaced on a spinner and rotated overnight. The reaction was thenfiltered by suction filtration and washed three times with CH₂Cl₂, threetimes with MeOH, once with 1:1 HOAc/CH₂Cl₂, 3 times with MeOH and thenlastly three times with CH₂Cl₂ to give R⁷SO₂-NR¹⁰CH₂CO—AA₂—AA₁-resin

[0230] 3b. For Z=—CO—:

[0231] To R¹⁰NHCH₂CO—AA₂-AA₁-resin is added sufficient CH₂Cl₂ is addedto swell the resin (˜12.5 mL/gr of resin). Three eq. of the desired acidchloride, R⁷COCl, and three eq. of Et₃N are then added. [A firstalternative is to add to the resin sufficient CH₂Cl₂ is added to swellthe resin (˜12.5 mL/gr of resin) followed by three eq. of the desiredcarboxylic acid, R⁷COOH, 3 eq. of diisopropylcarbodiimide and 0.05 eq.of a 0.116 M solution of dimethylaminopyridine in THF. A secondalternative is to add to the resin 3 eq. of the desired carboxylic acid,R⁷COOH, 3 eq. of HATU, 3 eq. HOAt and 6 eq. of DIPEA followed bysufficient NMP is added to swell the resin (˜12.5 mL/gr of resin ).] Thereaction was then placed on a spinner and rotated overnight. Thereaction was then filtered by suction filtration and washed three timeswith CH₂Cl₂, three times with MeOH, once with 1:1 HOAc/CH₂Cl₂, threetimes with MeOH and then lastly three times with CH₂Cl₂ to giveR⁷CO—NR¹⁰CH₂—AA₂—AA₁-resin

[0232] 3c. For Z=—CONH—:

[0233] R¹⁰NHCH₂CO—AA₂—AA₁-resin is first washed with THF, then 3 eq. ofthe desired isocyanate, R⁷N═C═O, was added followed by sufficient THF toswell the resin (˜12.5 mL/gr of resin). The reaction was then placed ona spinner and rotated overnight. The reaction was then filtered bysuction filtration and washed three times with CH₂Cl₂, three times withMeOH, once with 1:1 HOAc/CH₂Cl₂, three times with MeOH and then lastlythree times with CH₂Cl₂ to give R⁷NHCO—NR¹⁰CH₂—AA₂—AA₁-resin.

[0234] 4. Removal of Protecting Groups

[0235] If AA₁ or AA₂ contains an acid labile side chain protecting groupthat is to be removed, the resulting resin (R⁷Z—NR¹⁰CH₂CO—AA₂—AA₁-resin)was treated with a solution of 95/2.5/2.5 TFA/H₂O, triisopropyl silane(TIS) for 2 hours. The reaction was then filtered by suction filtrationand washed three times with CH₂Cl₂, three times with MeOH, and thenthree times with CH₂Cl₂.

[0236] 5. R⁷Z—NR¹⁰C(O)CH₂—AA₂—AA₁—NHOH

[0237] R⁷Z—NR¹⁰CH₂CO—AA₂—AA₁-resin was first washed with THF. SufficientTHF was added to swell the resin (˜12.5 mL/gr of resin) then 25 eq. of50% aq. NH₂OH was added and the reaction was rotated for two days. Thereaction was then filtered by suction filtration and washed with CH₂Cl₂,MeOH, and then CH₂Cl₂. The filtrate was concentrated under vacuum toobtain R⁷Z—NR¹⁰C(O)CH₂—AA₂—AA₁—NHOH

[0238] *Note in all cases, all solid reagents are added followed bysolvent. If the reagents are not solids, the solvent is added followedby the liquid reagents.

[0239] For AA₁ or AA₂ with side chain that require deprotection this wasdone just prior to the NH₂OH cleavage reaction. Typical amino acids withside chains requiring deprotection include Glu (OtBu), His (Boc) and Ser(OtBu).

Example XI Isolation and Preparation of Procollagen C-Proteinase

[0240] Cloning of Human PCP and Construction of the HT-1080 Vector

[0241] Human Procollagen C-Proteinase (PCP, also known as BoneMorphogenetic Protein-1 or BMP-1) was cloned by from a human fibroblastcDNA library (Stratagene, San Diego, Calif.). Cloning was performed byPCR based on the reported nucleotide sequence (Wozney, J. M., Rosen, V.,Celeste, A. J., Mitsock, L. M., Whitters, M. J., Kriz, R. W., Hewick, R.M., and Wang, E. A. (1989) direct GenBank submission accession M22488,locus HUMBMP1) using Taq polymerase, the 5′ primerGCGCGCGGTACCCGCCCCGCCAGCATGCCCGGCGTGGCCCGCCTGCCGCTGCTGCTCGGGCTGCTGCTGCTCCCGCGTCCCGGCCGGCCGCTGGACTTGGCCGACTACACCTAT GACCTGGC (SEQID NO:1)(Oligo Therapeutics, Inc., Wilsonville, Oreg.), and the 3′reverse strand primerCCGCTCGAGCCTCACTGGGGGGTCCGGTTTCTTTTCTGCACTCGGAATTTGAGCTGGT G (SEQ IDNO:2) (Gibco) to yield the entire full-length nucleotide encoding thesignal sequence, propeptide, catalytic domain, and all C-terminaldomains to the natural translation termination site. The PCR product waspurified by gel electrophoresis using the Wizard DNA Purification Kit(Promega, Madison, Wis.) and ligated directly into the mammalianexpression vector pCR3.1 (Invitrogen, Carlsbad, Calif.) by the TAcloning method. Ligated product was used to transform E coli strainTOP10F′ (Invitrogen, Carlsbad, Calif.) by a standard heat-shock method,and transformants were selected by restriction analysis of purifiedplasmid using the enzymes HindIII and BamHI. Transformants testingpositive for the PCP insert were submitted for sequencing using thePerkin-Elmer/ABI system. Two clones were selected that, combined,encoded the entire amino acid sequence identical to the one predicted byWozney et al. The two clones were recombined by restriction using theenzymes BbrI, which cleaved at a naturally occurring internal site, andEcoRV, which cleaved at the junction of the insert and vector. Theexcised fragments were religated into EcoRV-treated pCR3. 1. Theresulting construct contained the entire coding sequence identical tothat reported by Wozney et al. with the exception of two silentmutations in the signal sequence, G->A at both positions 39 and 45counting from the translation initiation site (ATG). The completedplasmid construct was amplified in E. coli DH5a and purified using anionexchange chromatography (MaxiPrep columns from Qiagen (Valencia, Calif.)catalog #12162).

[0242] Transfection of HT-1080 and Selection of the PCP-Expressing Clone

[0243] The human fibrosarcoma line HT-1080 (ATCC) was grown in highglucose DMEM (DMEM-HG) supplemented with 10% heat-inactivated fetalbovine serum (HI-FBS) in 100 mm culture dishes (Falcon, BectonDickenson, Franklin, N.J.) and transfected with 2 ug of purified plasmidusing the standard method for Lipofectamine (Gibco, Bethesda, Md.) inserum free medium. Stable transfectants were selected by treating theplated culture with 400 ug/ml G418 (Gibco). After selection for 10 days,the adherent single colonies were picked from the plate, replated in12-well plates and grown until confluent. Individual stable colonieswere screened for PCP expression by TaqMan (Perkin-Elmer, Foster City,Calif.) analysis using equivalent amounts of total RNA, the 5′ primerGACGAAGAGGACCTGAGGGCCTT (SEQ ID NO:3) (Perkin-Elmer, Foster City,Calif.), the 3′ reverse strand primer TTCCTGGAACTGCAGCTTTGA (SEQ IDNO:4) (Perkin-Elmer, Foster City, Calif.), and the reverse strand probeTGCCGTCTGAGATCCACAGCCTGCT (SEQ ID NO:5) (Perkin-Elmer). A stable line,HT-1080/hPCP-23, was chosen based on the highest PCP mRNA expressionlevel in the TaqMan screen. Stocks of the HT-1080/hPCP-23 stable linewere transferred to DMEM-HG supplemented with 5% HI-FBS and 10% DMSO (noG418 added) and were slowly frozen at −70° C. overnight, thentransferred to a liquid nitrogen bath for long-term storage. RevitalizedHT-1080/hPCP-23 were maintained in DMEM-HG supplemented with 10% HI-FBSand 250 ug/ml G418 for no more than 7 passages. Expression of PCP forharvest was carried out by replating and growing HT-1080/hPCP-23 on rattail type I collagen-coated plates (Falcon) in OptiMEM (Gibco) serumfree medium without G418 for 24 hr.

[0244] Production of PCP in HT 1080 Cells

[0245] The HT1080 cells that were transformed to produce PCP wereadapted to grow in suspension in optiMEM medium (GIBCO) supplementedwith 5% fetal bovine serum and 4 ml/L G418 (GIBCO). The culture wasmaintained at 37C and the dissolved oxygen at 30%. Typically batch sizesof 10 liters were produced. When the cell density reached 4-6×10⁵cells/ml, the culture fluid was collected and filtered through 0.2 ummembranes. Alternatively, the cell culture was perfused with fresh mediaat the rate of 0.8 to 1.0 culture volume/day. The density of theperfused cultures reached 1-2.5×10⁶ cells/ml and were maintained up totwo weeks with continuous harvests.

[0246] Purification of PCP from HT1080 Cells

[0247] A column packed with Dyematrex Gel Green A (Millipore, Bedford,Mass.) was equilibrated against 50 mM HEPES, pH 7.2, containing 6 mMCaCl₂ and 0.3M NaCl. After the HT1080 cell culture fluid was loaded, thecolumn was washed with 10 column volumes of the equilibration buffercontaining 1.0 M NaCl. PCP was eluted with 50 mM HEPES pH 7.2 containing3 M NaCl, 2 M urea and 6 mM CaCl₂. Eluate fractions were pooled andconcentrated to 150-200 mls and dialyzed against 4.0 liters of 50 mMHEPES, 6 mM CaCl₂, pH 7.2 overnight. The material was then centrifugedat 5,000 g for 15 minutes to remove precipitates. The PCP containingsample were stored at −20C until ready for further processing.

[0248] The PCP containing sample was thawed and diluted with 50 mM HEPESpH 7.2 containing 6 mM CaCl₂, if necessary to bring the NaClconcentration to 0.1-0.15 M The pH was adjusted to 6.7 with 2 N HCl. Theprotein solution was filtered through a 0.45 um filter to remove anyprecipitate. This preparation was then loaded onto a column packed withQ-Sepharose High Performance (Pharmacia, Piscataway, N.J.) which hadbeen equilibrated with 50 mM HEPES pH 6.7 containing 6 mM CaCl₂ and 0.15M NaCl. The PCP was not retained in the column and was therefore in theflow through fractions. The PCP was concentrated to 1 mg/ml and used forscreening.

[0249] Production of PCP in Drosophila Cells

[0250] Drosophila cells which had been transformed to produce PCP weregrown in bioreactors at a typical batch volume of 10 liters in SF900 IISF medium (GEBCO). The temperature was maintained at 30C and thedissolved oxygen at 30%. Periodically the cells were fed a cocktailconsisting of glutamine, lipids and yeastolate. When cell densitiesreached 30-50×10⁶ cells/ml, supernatants were harvested bycentrifugation and concentrated by ultrafiltration using a 30 Kdmembrane.

[0251] Purification of PCP from Culture Fluid from Drosophila Cells

[0252] Culture fluid from the drosophila cells was concentrated 8 foldand the pH adjusted to 7.1-7.2 if necessary. The culture fluid wascentrifuged at 3000 g for 10 minutes and filtered through 0.45 umfilters. The culture fluid was then loaded onto columns packed withcarboxy-sulfone packing material (J. T. Baker/Mallinckrodt,Phillipsburg, N.J.) which had been equilibrated with 0.1 M NaCl, 50 mMHEPES, 6 mM CaCl₂, pH 7.2. After being loaded, the column was washedwith 10 column volumes of the equilibration buffer. Retained proteinswere eluted with a gradient of 0.1 to 1.0 M NaCl in 9 column volumes.Fractions that had PCP activity were pooled for further purification.

[0253] The PCP eluted off the carboxy-sulfone column was loaded onto aDyematrex Gel Green A (Millipore, Bedford, Mass.) column that had beenequilibrated with 50 mM HEPES, pH 7.4 containing 0.3 M NaCl and 6 mMCaCl₂. The column was then washed with the equilibration buffercontaining 1 M NaCl. Retained proteins were eluted with 50 mM HEPES, pH7.4, 3 M NaCl, 2 M urea, 6 mM CaCl₂. The elution peak was concentratedand dialyzed against 50 mM HEPES, pH 7.4 containing 0.3 M NaCl, 6 mMCaCl₂. The preparation was centrifuged at 3000 g for 10 minutes. Brij 35(Sigma, Madison, Wis.) was added to the supernatant to a finalconcentration of 0.02%. This preparation was used for screening.

Example XII Isolation of Collagenase Enzymes

[0254] The catalytic domain of human collagenase-l was expressed as afusion protein with ubiquitin in E. Coli as described in Gehring, E. R.et al., J. Biol. Chem., 270, 22507, (1995). After purification of thefusion protein, the collagenase-1 catalytic domain was released bytreatment with 1 mM of aminophenylmercuric acetate (APMA) for 1 hour at37° C. and then purified by zinc chelate chromatography.

[0255] Human collagenase-2 and gelatinase B were isolated in active formfrom buffy coats as described in Mookhtiar, K. A. et al., Biochemistry,29, 10620, (1990).

[0256] The propeptide and catalytic domain portion of humancollagenase-3 was expressed in E. Coli as an N-terminal fusion proteinwith ubiquitin. After purification, the catalytic domain was released bytreatment with 1 mM APMA for 1 hour at 37° C., and then purified by zincchelate chromatography.

[0257] Rat collagenase-3 was purified in active form from the culturemedia of uterine smooth muscle cells as described in Roswit, W. T. etal., Arch. Biochem. Biophys., 225, 285-295 (1983).

Example XIII Inhibition of Procollagen C-Proteinase Activity

[0258] The ability of the compounds to inhibit PCP has been demonstratedin the following in vitro assays utilizing a synthetic peptide as thesubstrate.

[0259] Assay A

[0260] A continuous assay was performed using 20 μM substrate(Dabcyl-Pro-Tyr-Tyr-Gly-Asp-Glu-Pro-n-Leu-Edans) (SEQ ID NO:6). Thefinal assay conditions were 20 μM substrate, 50 mM HEPES pH 7.5, 50 mMNaCl, 3% DMSO, 37° C. and PCP enzyme. Product formation was monitored byfluorescence spectroscopy, Ex.=335 nm, Em.=490 nm. The IC₅₀ wascalculated from plots of the initial velocity vs. compoundconcentration.

[0261] Assay B

[0262] Eighty μL of buffer A (20 mM HEPES) containing the desiredconcentrations of the test compound in DMSO or carrier vehicle was mixedwith 10 L of approx. 1 mg/mL PCP enzyme and 10 L of 0.1 mM substrateboth in 20 mM HEPES. The contents are mixed, incubated at roomtemperature for 1-2 hours and fluorescent readings taken with a Victorplate reader (Ex. 405 nm, Em. 460 nM at 2000-40,000 lamp energy, 0.1-1sec/well). The substrate wasDACM-Cys-Pro-Tyr-Gly-Asp-Glu-Pro-nLeu-Lys-FITC-OH. (SEQ ID NO:7)(DACM=dimethylaminocoumarylmaleimide, FITC=fluorescein isothiocyanate).The IC₅₀ was calculated from plots of the initial velocity vs. compoundconcentration.

[0263] Additional in vitro assays using native procollagen as thesubstrate may also be used and these assays are described in more detailin WO 97/05865 (“C-Proteinase Inhibitors for the Treatment of DisordersRelating to the Overproduction of Collagen”), published Feb. 20, 1997.

[0264] The compounds in Tables I-III, V and VI had IC₅₀'s in the rangeof 0.02 to 200 μM.

[0265] The compounds in Table IV had IC₅₀'s in the range of 10-1000 μM.

Example XIV Measurement of Collagenase Activity

[0266] The collagenase-1, collagenase-2 and collagenase-3 inhibitoryactivity of compounds of this invention in vitro was determined based onthe hydrolysis of MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂ (SEQ ID NO:8)(Bachem, Inc.) at 37° C. as described in Knight, C. G., et al., FEBSLett., 296(3): 263-266 (1992).

[0267] The collagenase enzyme was diluted with assay buffer (50 mMTricine pH 7.5, 200 mM NaCl, 10 mM CaCl₂, and 0.005% Brij-35) containing10 gmole of above substrate dissolved in DMSO. Compounds of theinvention dissolved in DMSO or only DMSO (control samples) were addedsuch that the final DMSO concentration in all assays was 2.5%. Thefluorescence changes were monitored with a Perkin-Elmer LS-50Bfluorimeter using an excitation wavelength of 328 nm and an emissionwavelength of 393 nm.

[0268] Selected compounds from Tables I-VI were 10-1000 more selectivefor PCP inhibition than for the human collagenase enzymes.

[0269] The foregoing invention has been described in some detail by wayof illustration and example, for purposes of clarity and understanding.It will be obvious to one of skill in the art that changes andmodifications may be practiced within the scope of the appended claims.Therefore, it is to be understood that the above description is intendedto be illustrative and not restrictive. The scope of the inventionshould, therefore, be determined not with reference to the abovedescription, but should instead be determined with reference to thefollowing appended claims, along with the full scope of equivalents towhich such claims are entitled.

[0270] All patents, patent applications and publications cited in thisapplication are hereby incorporated by reference in their entirety forall purposes to the same extent as if each individual patent, patentapplication or publication were so individually denoted.

1 8 1 122 DNA Artificial Sequence pcr primer 1 gcgcgcggta cccgccccgccagcatgccc ggcgtggccc gcctgccgct gctgctcggg 60 ctgctgctgc tcccgcgtcccggccggccg ctggacttgg ccgactacac ctatgacctg 120 gc 122 2 59 DNAArtificial Sequence pcr primer 2 ccgctcgagc ctcactgggg ggtccggtttcttttctgca ctcggaattt gagctggtg 59 3 23 DNA Artificial Sequence pcrprimer 3 gacgaagagg acctgagggc ctt 23 4 21 DNA Artificial Sequence pcrprimer 4 ttcctggaac tgcagctttg a 21 5 25 DNA Artificial Sequence pcrprimer 5 tgccgtctga gatccacagc ctgct 25 6 8 PRT Artificial Sequencesynthetic peptide 6 Pro Tyr Tyr Gly Asp Glu Pro Leu 1 5 7 9 PRTArtificial Sequence synthetic peptide 7 Cys Pro Tyr Gly Asp Glu Pro LeuLys 1 5 8 7 PRT Artificial Sequence synthetic peptide 8 Arg Ala Xaa LeuGly Leu Pro 1 5

What is claimed is:
 1. A compound selected from the group of compoundsrepresented by Formula (I):

wherein: R¹ and R⁴ are, independently of each other, hydrogen or alkyl;R² is: (i) cycloalkyl, cycloalkylalkyl, aryl, aralkyl, aralkenyl,heteroaryl, heteroaralkyl, heteroaralkenyl, heterocyclo orheterocycloalkyl; or (ii) -(alkylene)-B—X where B is —O—, —NR⁸—,—S(O)_(n)— (where n is 0, 1 or 2), —C═O, —CONR⁸—, —NR⁸CO₂—, NR⁸SO₂— or—C(=NR⁸)NR⁸SO₂— (where R⁸ is H or alkyl), and X is cycloalkyl,cycloalkylalkyl, aryl, aralkyl heteroaryl or heteroaralkyl; or (iii)-(alkylene)-B—X where B is —NR⁸CO— (where R⁸ is H or alkyl), and X iscycloalkyl, cycloalkylalkyl, aryl, aralkyl heteroaryl or heteroaralkyl;or (iv) R² and R³ form an alkylene or heteroalkylene chain; R³ ishydrogen or alkyl; R⁶ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl,aryl, aralkyl, heteroaryl or heteroaralkyl; R⁵ is: (i) hydrogen, alkyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, aralkenyl, heteroaryl,heteroaralkyl, heteroaralkenyl, heterocycloalkyl, heteroalkyl, or-(alkylene)-C(O)—X¹ where X¹ is alkyl, hydroxy, alkoxy, aryl, aralkyl,aryloxy, aralkyloxy, heteroaryl, heteroaryloxy, heteroaralkyloxy orNR′R″ (where R′ and R″ are independently H or alkyl, or R′ and R″ forman alkylene chain); or (ii) R⁵ and R⁴ form an alkylene chain; or (iii)R⁵ and R⁶ form an alkylene chain; n is 0 or 1; A is—C(═O)—CH(R⁹)—(CH₂)_(m)—N(R¹⁰)— wherein: m is an integer from 0-5inclusive; R⁹ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl,aralkyl, heteroaryl, heteroaralkyl, heterocycloalkyl, heteroalkyl, or-(alkylene)-C(O)—X¹ where X¹ is alkyl, hydroxy, alkoxy, aryl, aralkyl,aryloxy, aralkyloxy, heteroaryl, heteroaryloxy, heteroaralkyloxy orNR′R″ (where R′ and R″ are independently H or alkyl, or R′ and R″ forman alkylene chain); and R¹⁰ is hydrogen, alkyl, aralkyl orheteroaralkyl; Z is Y—B wherein: Y is alkylene or a bond; and B is —CO—,—C(O)O—, —CONR⁸—, —SO₂—, or —SO₂NR⁸— (where R⁸ is hydrogen or alkyl),alkylene (optionally substituted by hydroxy, alkoxy, amino,monoalkylamino or dialkylamino) or a bond; R⁷ is cycloalkyl,cycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroaralkyl; providedthat when n=0 and Z is SO₂, then R² does not contain an imidazole group;and their pharmaceutically acceptable salts, prodrugs, individualisomers, and mixtures of isomers.
 2. The compound of claim 1 wherein: nis
 0. 3. The compound of claim 2 wherein R³and R⁶ are hydrogen.
 4. Thecompound of claim 3, wherein: R²is aralkyl or heteroaralkyl.
 5. Thecompound of claim 4 wherein: Z is —C(O)O— or —S(O)₂—.
 6. The compound ofclaim 5 wherein: R² is optionally substituted benzyl orheteroaralkylmethyl.
 7. The compound of claim 6 wherein, R² is4-t-butoxybenzyl, 3-chlorobenzyl, 3-indolyl methyl, 2-thienylmethyl,4-imidazolylmethyl or 4-thiazolylmethyl.
 8. The compound of claim 7wherein R² is 4-thiazolylmethyl.
 9. The compound of claim 8 wherein: R⁷is aryl, aralkyl, heteroaryl or heteroaralkyl.
 10. The compound of claim8 wherein: Z is —C(O)O— and R⁷ is optionally substituted benzyl.
 11. Thecompound of claim 9 wherein: Z is —SO₂— and R⁷ is aryl or heteroaryl.12. The compound of claim 10, wherein: R¹ and R⁴ are hydrogen and R⁵ isalkyl.
 13. The compound of claim 12 wherein R⁵ is (S,S)-1-methylpropyl.14. The compound of claim 11, wherein: R¹ and R⁴ are hydrogen and R⁵ isalkyl.
 15. The compound of claim 14 wherein R⁵ is (S,S)-1-methylpropyl.16. The compound of claim 3, wherein: R² is (alkylene)-B—X where B is—O—, —NR⁸—, —S(O)_(n)— (where n is 0, 1 or 2), —C═O, —CONR⁸—, —NR⁸CO₂—,—NR⁸SO₂— or —C(═NR⁸)NSO₂— (where R⁸ is H or alkyl), and X is cycloalkyl,cycloalkylalkyl, aryl, aralkyl heteroaryl or heteroaralkyl.
 17. Thecompound of claim 16, wherein: Z is —C(O)O— or —S(O)₂—.
 18. The compoundof claim 17, wherein R² is CH₂—B—X and B is —NHCO₂— and X is benzyl. 19.The compound of claim 18 wherein: R⁷ is aryl or aralkyl.
 20. Thecompound of claim 19, wherein: R¹ and R⁴ are hydrogen and R⁵ is alkyl.21. The compound of claim 20 wherein R⁵ is (S,S)-1-methylpropyl.
 22. Thecompound of claim 1 wherein: n is
 1. 23. The compound of claim 22wherein m is 0 and R³ and R⁶ are hydrogen.
 24. The compound of claim 23,wherein: R² is aralkyl or heteroaralkyl.
 25. The compound of claim 24,wherein: Z is —C(O)O— or —S(O)₂—.
 26. The compound of claim 25, wherein:R² is optionally substituted benzyl or heteroarylmethyl.
 27. Thecompound of claim 26 wherein R is 4-t-butoxybenzyl, 3-chlorobenzyl,3-indolyl methyl, 2-thienylmethyl, 4-imidazolylmethyl or4-thiazolylmethyl.
 28. The compound of claim 27 wherein R² is4-thiazolylmethyl.
 29. The compound of claim 28 wherein: R⁷ is aryl,aralkyl, heteroaryl or heteroaralkyl.
 30. The compound of claim 29wherein: Z is —C(O)O— and R⁷ is benzyl.
 31. The compound of claim 29wherein: Z is —SO₂— and R⁷ is aryl.
 32. The compound of claim 30,wherein: R¹ and R⁴ are hydrogen and R⁵ is alkyl.
 33. The compound ofclaim 32 wherein R⁵ is (S,S)-1-methylpropyl.
 34. The compound of claim31, wherein: R¹ and R⁴ are hydrogen and R⁵ is alkyl.
 35. The compound ofclaim 34 wherein R⁵ is (S,S)-1-methylpropyl.
 36. The compound of claim23, wherein: R² is (alkylene)-B—X where B is —O—, —NR⁸—, —S—, —C═O,—CONR⁸—, —NR⁸CO₂—, —NSO₂— or —C(═NR⁸)NSO₂— (where R⁸ is H or alkyl), andX is cycloalkyl, cycloalkylalkyl, aryl, aralkyl heteroaryl orheteroaralkyl.
 37. The compound of claim 36, wherein: Z is —C(O)O— or—S(O)₂—.
 38. The compound of claim 37, wherein R² is CH₂—B—X and B is—NHCO₂— and X is benzyl.
 39. The compound of claim 38 wherein: R⁷ isaryl or aralkyl.
 40. The compound of claim 39, wherein: R¹ and R⁴ arehydrogen and R⁵ is alkyl.
 41. The compound of claim 40 wherein R⁵ is(S,S)-1-methylpropyl.
 42. A pharmaceutical composition comprising thecompound of claim 1 and a pharmaceutically acceptable excipient.
 43. Amethod of treating disease comprising administering to a patient in needthereof a compound of claim
 1. 44. The method of claim 43, wherein thedisease is a fibrotic disease.
 45. The method of claim 44 wherein thedisease is acute respiratory distress syndrome.
 46. A method of treatingfibrosis comprising administering to a patient in need thereof aninhibitor of procollagen C-proteinase that is at least ten-fold moreselective for procollagen C-proteinase over both collagenase-1,collagenase-2 and collagenase-3.
 47. A method for preparing thecompounds of claim 1 by: (i) treating a compound of Formula II wherein Lis a leaving group and R¹-R⁷, A, n and Z are as defined in claim 1 withhydroxylamine or a protected derivative thereof, and (ii) deprotectingas necessary and isolating the compound of claim 1.